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Epigenetics is increasingly recognized as an important molecular mechanism underlying phenotypic variation. To study DNA methylation in ecological and evolutionary contexts, epiRADseq is a cost‐effective next‐generation sequencing (NGS) technique based on reduced representation sequencing of genomic regions surrounding non‐/methylated sites. EpiRADseq for genome‐wide methylation abundance and ddRADseq for genome‐wide single‐nucleotide polymorphism (SNP) genotyping follow very similar library and sequencing protocols, but to date these two types of dataset have been handled separately. Here we test the performance of using epiRADseq data to generate SNPs for population genomic analyses.We tested the robustness of using epiRADseq data for population genomics with two independent datasets: a newly generated single‐end dataset for the European whitefish Coregonus lavaretus, and a re‐analysis of publicly available, previously published paired‐end data on corals. Using standard bioinformatic pipelines with a reference genome and without (i.e. de novo catalogue loci), we compared the number of SNPs retained, population genetic summary statistics and population genetic structure between data drawn from ddRADseq and epiRADseq library preparations.We found that SNPs drawn from epiRADseq are similar in number to those drawn from ddRADseq, with 55%–83% of SNPs being identified by both methods. Genotyping error rate was <5% in both approaches. EpiRADseq‐specific allele dropout was low (~1%). For summary statistics, such as heterozygosity and nucleotide diversity, there is a strong correlation between methods (Spearman's rho > 0.88). Furthermore, identical patterns of population genetic structure were recovered using SNPs from epiRADseq and ddRADseq approaches.We show that SNPs obtained from epiRADseq are highly similar to those from ddRADseq and are equivalent for estimating genetic diversity and population structure. This finding is particularly relevant to researchers interested in genetics and epigenetics on the same individuals because using a single epigenomic approach to generate two datasets greatly reduces the time and financial costs compared to using these techniques separately. It also efficiently enables correction of epigenetic estimates with population genetic data. Many studies will benefit from a combinatorial approach with genetic and epigenetic markers and this demonstrates a single, efficient method to do so. [ABSTRACT FROM AUTHOR]

Background. Microsatellite loci, or single sequence repeats (SSR), are widely used as powerful markers in population genetics. They represent an attractive tool for studying plants such as grasses, whose evolution is driven by hybridisation and polyploidization. However, the development of microsatellite markers has been challenging and time-consuming, especially for non-model organisms lacking available genome-wide sequence data. One straightforward and low-cost approach is to transfer the SSR loci developed for one species, or complex, to another closely-related one. This work evaluates the transferability of microsatellite loci from homoploid to allopolyploid complexes of fine-leaved Festuca species and to assess their use in two new species. The studied complex (F. amethystina-F. tatrae) is a useful model for research on the local adaptability of grasses with different ploidy levels. Since both species can be considered as rare or threatened (F. tatrae-as a mountain and narrow endemic species and F. amethystina-a mountain species with relict lowland populations), any tool enabling studies on genetic diversity and population genetics, such as SSR markers, could also be very useful in a conservation context. Methods. The ploidy level within populations was estimated using flow cytometry. One diploid and one tetraploid population of F. amethystina and a diploid population of F. tatrae were chosen to test the transferability of SSR loci. Because our work describes the transfer of SSR nuclear markers designed originally for F. gautieri, a phylogenetic tree was prepared based on the ITS marker to assess the genetic distance between the studied complexes. The PCR products were separated on a high-resolution agarose gel, intended for SSR marker analysis. Appropriate solutions for the allotetraploid population and whole mixed-ploidy complex were implemented. Results. Flow cytometry confirmed earlier data regarding DNA content in the investigated species and cytotypes. The phylogenetic ITS tree indicated a small genetic distance between F. gautieri complexes and the studied species. Ten microsatellite markers were successfully transferred. All markers were polymorphic. In total, 163 different alleles were scored from the 10 SSR loci. PCoA of accessions revealed well-separated groups corresponding to studied populations. Over 60% of the total variance is explained by differentiation within populations and one third among them. Conclusions. The transferred markers are valid tools for the study of population genetics and inheritance relationships within cytotypes and species and between them. The presented markers can be used to study inbreeding depression in the Festuca species, and variations in the degrees of genetic diversity between different cytotypes in mountain and lowland areas. Our findings can also be applied to study conservation strategies for ensuring biodiversity at the genetic level in polyploid complexes. [ABSTRACT FROM AUTHOR]

A decline in soil biodiversity is generally considered to be the reduction of forms of life living in soils, both in terms of quantity and variety. Where soil biodiversity decline occurs, it can significantly affect the soils' ability to function, respond to perturbations and recover from a disturbance. Several soil threats have been identified as having negative effects on soil biodiversity, including human intensive exploitation, land-use change and soil organic matter decline. In this review we consider what we mean by soil biodiversity, and why it is important to monitor. After a thorough review of the literature identified on a Web of Science search concerning threats to soil biodiversity (topic search: threat* "soil biodiversity''), we compiled a table of biodiversity threats considered in each paper including climate change, land use change, intensive human exploitation, decline in soil health or plastic; followed by detailed listings of threats studied. This we compared to a previously published expert assessment of threats to soil biodiversity. In addition, we identified emerging threats, particularly microplastics, in the 10 years following these knowledge based rankings. We found that many soil biodiversity studies do not focus on biodiversity sensu stricto, rather these studies examined either changes in abundance and/or diversity of individual groups of soil biota, instead of soil biodiversity as a whole, encompassing all levels of the soil food web. This highlights the complexity of soil biodiversity which is often impractical to assess in all but the largest studies. Published global scientific activity was only partially related to the threats identified by the expert panel assessment. The number of threats and the priority given to the threats (by number of publications) were quite different, indicating a disparity between research actions versus perceived threats. The lack of research effort in key areas of high priority in the threats to soil biodiversity are a concerning finding and requires some consideration and debate in the research community. [ABSTRACT FROM AUTHOR]

Exceptionally well-preserved fossils are frequently encased by carbonate concretions. The initial steps of their formation in marine and freshwater sediments are induced by microbial activity. The role of the involved microbial communities, however, is not well understood. In this study, siderite (FeCO3) formation in microbial microcosms is observed, with various fatty acyl compounds (lipids, surfactants) as substrates and Wadden Sea sediment samples as inocula. In actively growing microcosms, sulfate-reducing bacteria (the genus Desulfofrigus in particular) dominate the microbial community and submicroscopic siderite precipitates on bacterial cell surfaces were identified. We suggest that these biologically induced mineralization processes may, in the natural environment, initiate the formation of large concretions under suboxic conditions in coastal sediments. [ABSTRACT FROM AUTHOR]

The development of agro-ecosystems in the pastures of the Pampean Region has substantially modified their structure and functioning. Many wild mammal populations in the Argentinean Pampas face habitat loss and/or fragmentation due to human activities, resulting in harmful genetic effects. The screaming hairy armadillo (Chaetophractus vellerosus) is a species considered an indicator of the state of preservation of the environments it inhabits. However, very little information is available about its mating system in the wild. In this sense, an isolated population of the screaming hairy armadillo in the northeast of Buenos Aires Province, which is separated from the main distribution area of the species by about 500 km, requires special attention. Genetic studies that analyzed social behavior and mating systems in Xenarthra are scarce but necessary to establish conservation actions for the isolated screaming hairy armadillo population under study. Thus, we analyzed the existence of a possible social organization in the species, together with its mating system, using a set of previously characterized microsatellites. Our results showed a complex scenario for the dispersal and mating system in this C. vellerosus population. Males disperse and females have a philopatric tendency with some degree of dispersal. This strategy, in combination with a polygynous-polyandrous mating system, could enhance genetic variability in this small and isolated population. In addition, no evidence of social organization was found. [ABSTRACT FROM AUTHOR]

Reproduction and recruitment are essential processes for the continued success of coral communities. Islas Marias Archipelago is considered a connectivity node among coral communities distributed along the Northeastern Tropical Pacific (NTP); as such, sexual reproduction of scleractinian corals affects the maintenance of the local populations and the long-distance connectivity of reefs across the region. In this study, successful sexual reproduction in the species of the three most abundant scleractinian corals genera was demonstrated, in part by documenting gamete presence and maturation in tissues and spatio-temporal variability in juvenile coral settlement, which were quantified across substrate type, habitat quality, and environmental factors. Only 12 larvae recruited (ten Porites, two Pavona) to artificial substrates, and monthly recruitment density of 1.82 ± 0.23 recruits m−2 (n = 383) was recorded on natural substrates. There were significant differences between genera, with Porites producing the highest density of recruits (0.60 ± 0.45 recruits m−2, followed by Pavona (0.52 ± 0.24 recruits m−2) and Pocillopora (0.28 ± 0.06 recruits m−2). The highest coral recruitment was observed at Baby Reef (2.57 ± 0.46 recruits m−2), followed by Cleofas II (1.81 ± 0.21 recruits m−2) and Japanese Garden (1.24 ± 0.46 recruits m−2); the former site was characterized by the highest cover of crustose coralline algae. We found successful recruitment of the main reef-forming species in the Mexican Pacific and suggest that Islas Marias is a region of critical importance in terms of its function as a source of genetic variability, and, the generation of, new individuals that will be key to sustaining coral reef ecosystems in the NTP. [ABSTRACT FROM AUTHOR]

Reclamation of anthropogenically impacted environments is a critical issue worldwide. In the oil sands extraction industry of Alberta, reclamation of mining‐impacted areas, especially areas affected by tailings waste, is an important aspect of the mining life cycle. A reclamation technique currently under study is water‐capping, where tailings are capped by water to create an end‐pit lake (EPL). Base Mine Lake (BML) is the first full‐scale end‐pit lake in the Alberta oil sands region. In this study, we sequenced eukaryotic 18S rRNA genes recovered from 92 samples of Base Mine Lake water in a comprehensive sampling programme covering the ice‐free period of 2015. The 565 operational taxonomic units (OTUs) generated revealed a dynamic and diverse community including abundant Microsporidia, Ciliata and Cercozoa, though 41% of OTUs were not classifiable below the phylum level by comparison to 18S rRNA databases. Phylogenetic analysis of five heterotrophic phyla (Cercozoa, Fungi, Ciliata, Amoebozoa and Excavata) revealed substantial novel diversity, with many clusters of OTUs that were more similar to each other than to any reference sequence. All of these groups are entirely or mostly heterotrophic, as a relatively small number of definitively photosynthetic clades were amplified from the BML samples. [ABSTRACT FROM AUTHOR]

The choice of sampling method is essential in the development of a population genetic study. This choice influences the type of samples that can be obtained and, therefore, the quality of the extracted DNA. Sampling methods can be classified into invasive, non-invasive, and minimally invasive. This last category is preferred, as it minimizes handling, damage, and degree of disturbance to animals, while still providing enough DNA for genetic studies. Here, we describe a very low cost and easy to manufacture minimally invasive sampler, developed to obtain hair samples from pinnipeds. The sampler was highly effective both at reaching animals and collecting a good number of hairs. We compared the efficiency of hair samples obtained using our device, versus tissue samples collected from dead animals, by counting successful mtDNA amplifications and sequencing. Hair samples amplified more efficiently compared with tissue samples, but sequencing was equally efficient for both sample types. In conclusion, the sampler described is an excellent option to obtain DNA for genetic studies. This minimally invasive technique can also be used to sample hair for other studies, such as estimation of environmental contaminants, measurement of individual hormone levels, or diet determination through the analysis of stable isotopes, among other potential uses. [ABSTRACT FROM AUTHOR]

While metabarcoding and metagenomic approaches are increasingly popular, questions remain about how best to analyse and taxonomically characterize the sequence data produced by such methods. Due to a lack of software infrastructure, important reference sequence curation steps are often ignored.We present MetaCurator, a software package designed for automated reference sequence curation and highly generalizable across markers and study systems. MetaCurator contains two signature tools. IterRazor utilizes profile hidden Markov models and an iterative search framework to exhaustively identify and extract the precise marker of interest from available references. DerepByTaxonomy dereplicates sequences using a taxonomically aware approach, removing duplicates only when they belong to the same taxon. This is important for highly conserved markers, such as plant rbcL and trnL, which often display no sequence divergence across taxa, even at the genus level.Using MetaCurator, we produced reference sequence databases for a popular arthropod COI marker as well as four plant barcoding markers, trnL, rbcL, ITS2 and trnH. In comparing these databases to those produced by recent and comparable studies, we show that the Metacurator pipeline exhibits greater sensitivity during sequence extraction, especially for poorly conserved markers. Further, database taxonomic richness was not decreased following sequence dereplication, as observed in previous studies.MetaCurator is supported on OSX and Linux and is freely available under a GPL v3.0 license at https://github.com/RTRichar/MetaCurator. The reference databases produced in this work, and the commands used for curation, are available at https://github.com/RTRichar/MetabarcodeDBsV2. [ABSTRACT FROM AUTHOR]

An extraordinarily high intraspecific chemical diversity, i.e. chemodiversity, has been found in several plant species, of which some are of major ecological or economic relevance. Moreover, even within an individual plant there is substantial chemodiversity among tissues and across seasons. This chemodiversity likely has pronounced ecological effects on plant mutualists and antagonists, associated foodwebs and, ultimately, biodiversity. Surprisingly, studies on interactions between plants and their herbivores or pollinators often neglect plant chemistry as a level of diversity and phenotypic variation. The main aim of this Research Unit (RU) is to understand the emergence and maintenance of intraspecific chemodiversity in plants. We address the following central questions: 1) How does plant chemodiversity vary across levels, i.e., within individuals, among individuals within populations, and among populations? 2) What are the ecological consequences of intraspecific plant chemodiversity? 3) How is plant chemodiversity genetically determined and maintained? By combining field and laboratory studies with metabolomics, transcriptomics, genetic tools, statistical data analysis and modelling, we aim to understand causes and consequences of plant chemodiversity and elucidate its impacts on the interactions of plants with their biotic environment. Furthermore, we want to identify general principles, which hold across different species, and develop meaningful measures to describe the fascinating diversity of defence chemicals in plants. These tasks require integrated scientific collaboration of experts in experimental and theoretical ecology, including chemical and molecular ecology, (bio)chemistry and evolution. [ABSTRACT FROM AUTHOR]

A detailed study is carried out of the accuracy of molecular equilibrium geometries obtained from least-squares fits involving experimental rotational constants B[sub 0] and sums of ab initio vibration–rotation interaction constants α[sub r][sup B]. The vibration–rotation interaction constants have been calculated for 18 single-configuration dominated molecules containing hydrogen and first-row atoms at various standard levels of ab initio theory. Comparisons with the experimental data and tests for the internal consistency of the calculations show that the equilibrium structures generated using Hartree–Fock vibration–rotation interaction constants have an accuracy similar to that obtained by a direct minimization of the CCSD(T) energy. The most accurate vibration–rotation interaction constants are those calculated at the CCSD(T)/cc-pVQZ level. The equilibrium bond distances determined from these interaction constants have relative errors of 0.02%–0.06%, surpassing the accuracy obtainable either by purely experimental techniques (except for the smallest systems such as diatomics) or by ab initio methods. © 2002 American Institute of Physics. [ABSTRACT FROM AUTHOR]

Recently developed non-invasive environmental DNA-based (eDNA) techniques have enlightened modern conservation biology, propelling the monitoring/management of natural populations to a more effective and efficient approach, compared to traditional surveys. However, due to rapid-expansion of eDNA, confusion in terminology and collection/analytical pipelines can potentially jeopardize research progression, methodological standardization, and practitioner adoption in several ways. Present investigation reflects the developmental progress of eDNA (sensu stricto) including highlighting the successful case studies in conservation management. The eDNA technique is successfully relevant in several areas of conservation research (invasive/conserve species detection) with a high accuracy and authentication, which gradually upgrading modern conservation approaches. The eDNA technique related bioinformatics (e.g., taxon-specific-primers MiFish, MiBird, etc.), sample-dependent methodology, and advancement of sequencing technology (e.g., oxford-nanopore-sequencing) are helping in research progress. The investigation shows that the eDNA technique is applicable largely in (i) early detection of invasive species, (ii) species detection for conservation, (iii) community level biodiversity monitoring, (iv) ecosystem health monitoring, (v) study on trophic interactions, etc. Thus, the eDNA technique with a high accuracy and authentication can be applicable alone or coupled with traditional surveys in conservation biology. However, a comprehensive eDNA-based monitoring program (ecosystem modeling and function) is essential on a global scale for future management decisions.

This paper showcases the development of plant virology in Croatia at the University of Zagreb, Faculty of Science, from its beginning in the 1950s until today, more than 70 years later. The main achievements of the previous and current group members are highlighted according to various research topics and fields. Expectedly, some of those accomplishments remained within the field of plant virology, but others make part of a much-extended research spectrum exploring subviral pathogens, prokaryotic plant pathogens, fungi and their viruses, as well as their interactions within ecosystems. Thus, the legacy of plant virology in Croatia continues to contribute to the state of the art of microbiology far beyond virology. Research problems pertinent for directing the future research endeavors are also proposed in this review.

High-throughput molecular methods such as quantitative polymerase chain reaction (qPCR) and environmental microarrays are cost-effective methods for semi-quantitative assessment of bacterial community structure and the identification of specific target organisms. Both techniques rely on short nucleotide sequences, so-called oligonucleotide probes, which require high specificity to the organisms in question to avoid cross-hybridization with non-target taxa. However, designing oligonucleotide probes for novel taxa or marker genes that show sufficient phylogenetic sensitivity and specificity is often time- and labor-intensive, as each probe has to be in-silico tested for its specificity and sensitivity. Here we present ProbeSpec, to our knowledge the first batch sensitivity and specificity estimation and visualization tool for oligonucleotide probes integrated into the widely used ARB software. Using ProbeSpec’s interactive “mismatch threshold” and “clade marked threshold” we were able to reduce the development time of highly specific probes for a recently published environmental oligonucleotide microarray from several months to one week.

The increasing demand for ornamental, cosmetic and pharmaceutical products is driving exploitation of plant species globally. Sub-Saharan Africa harbours unique and valuable plant resources and is now a target of plant resource depletion. African Sandalwood (Osyris lanceolata), a multi-purpose and drought-tolerant species, has seen increased exploitation for the last thirty years and is now declared endangered. Initiatives to conserve O. lanceolata are not yet successful in Africa due to poor understanding of the species. This review surveys relevant research on the ecology, taxonomy, population dynamics, genetic diversity and ethnobotany of O. lanceolata, and highlights gaps in the literature for further research. A scoping review of grey literature, scholarly papers and reports was applied with pre-determined criteria to screen relevant information. Review findings indicate O. lanceolata is a globally distributed species with no identified center of origin. In Africa, it ranges from Algeria to Ethiopia and south to South Africa; in Europe it occurs in the Iberian Peninsula and Balearic Islands; in Asia from India to China, and also on Socotra. The species has a confusing taxonomy, with unresolved issues in nomenclature, country range distribution, extensive synonymisation and variation in growth form (shrub or tree). The species population is reported to be declining in Africa, but information on population dynamics across its entire range of distribution is anecdotal. Additionally, ecological factors influencing spatial distribution and survival of the species remain unknown. A variety of uses are reported for O. lanceolata globally, including: cultural; medicinal and food; dye; perfumery; timber; ethnoveterinary and phytoremediation. Key research areas and implications for conservation of O. lanceolata in Sub-Saharan Africa are proposed.

Intracellular bacteria of the genus Wolbachia (α-Proteobacteria) are the most widespread endosymbionts of insects. Host infection is usually associated with alterations in reproduction, such as cytoplasmic incompatibility, the induction of parthenogenesis and offspring sex ratio bias: all phenomena that may influence host speciation. In the present study, by using well-established molecular tools, we investigated the presence of Wolbachia in leaf beetles of the genus Crioceris and their host plants, which are various species of Asparagus. Multilocus sequence typing of bacterial genes showed that despite their occurrence in the same habitat and feeding on the same plant, two species of Crioceris, C. quinquepunctata and C. quatuordecimpunctata, are infected by two different strains of Wolbachia. C. asparagi, C. paracenthesis and C. duodecimpunctata, which are sympatric with the infected species, do not harbour the bacterium. Interestingly, DNA of Wolbachia was detected in host plant tissues that are exploited by the beetles, providing evidence for the horizontal transmission of the bacterium between beetles and their host plants. Moreover, Wolbachia was detected in species of Crioceris that are not closely related.

Microcins are low-molecular-weight, ribosomally produced, highly stable, bacterial-inhibitory molecules involved in competitive, and amensalistic interactions between Enterobacteriaceae in the intestine. These interactions take place in a highly complex chemical landscape, the intestinal eco-active chemosphere, composed of chemical substances that positively or negatively influence bacterial growth, including those originated from nutrient uptake, and those produced by the action of the human or animal host and the intestinal microbiome. The contribution of bacteria results from their effect on the host generated molecules, on food and digested food, and organic substances from microbial origin, including from bacterial degradation. Here, we comprehensively review the main chemical substances present in the human intestinal chemosphere, particularly of those having inhibitory effects on microorganisms. With this background, and focusing on Enterobacteriaceae, the most relevant human pathogens from the intestinal microbiota, the microcin’s history and classification, mechanisms of action, and mechanisms involved in microcin’s immunity (in microcin producers) and resistance (non-producers) are reviewed. Products from the chemosphere likely modulate the ecological effects of microcin activity. Several cross-resistance mechanisms are shared by microcins, colicins, bacteriophages, and some conventional antibiotics, which are expected to produce cross-effects. Double-microcin-producing strains (such as microcins MccM and MccH47) have been successfully used for decades in the control of pathogenic gut organisms. Microcins are associated with successful gut colonization, facilitating translocation and invasion, leading to bacteremia, and urinary tract infections. In fact, Escherichia coli strains from the more invasive phylogroups (e.g., B2) are frequently microcinogenic. A publicly accessible APD3 database http://aps.unmc.edu/AP/ shows particular genes encoding microcins in 34.1% of E. coli strains (mostly MccV, MccM, MccH47, and MccI47), and much less in Shigella and Salmonella (

Studies on the population structure of common widespread macroalgae in the Gulf of Mexico (GoMx) are scarce, and this knowledge gap limits our understanding on how disturbances affect the genetic diversity of macroalgae in this basin. The latter is due to the lack of a baseline that can be compared with allele frequency surveys conducted after a major disturbance such as the 2010 Deepwater Horizon oil spill (DWH), which leaked 780,000 m3 of crude oil in the vicinity of highly diverse macroalgal communities. Fortunately, quantitative assessments of the population structure pre-DWH can be accomplished for several macroalgae with dried specimens collected from research cruises conducted before 2010 in the offshore GoMx. Based on three markers (cytochrome c oxidase subunit I, COX II-III intergenic spacer, and the RuBisCO large subunit), this study reconstructed the allele frequencies pre-DWH for a GoMx-widespread macroalga, Botryocladia occidentalis, and revealed the existence of distinct populations in each of three distant regions of the GoMx: Florida Middle Grounds (FL), Campeche Banks (CB), and offshore Louisiana (LA). Population structure was assessed with exact tests of population differentiation and Analyses of Molecular Variance. FL harbored the most differentiated and genetically diverse population due to the presence and abundance of unique haplotypes. Interestingly, FL haplotypes were not closely phylogenetically related to each other and included the most divergent lineages of the entire GoMx; this phylogeographic pattern suggests a strong influence of migrants from the Caribbean on the FL population. Additionally, likelihood ratio tests with a small sample collected post-DWH indicated that the LA population underwent strong changes, showing statistically significant differences before (LA) vs. after (L2) the disaster. Whereas the LA population had affinity to CB, L2 showed a FL haplotype that, before the disaster, had never been reported in LA or CB. Such changes may not be permanent but rather a temporary response to disturbance; also, they may not necessarily be caused by the spilled oil but by other factors associated with the DWH.

The western Palaearctic species of the hawkmoth genus Hyles (Lepidoptera: Sphingidae) have long been the subject of molecular phylogenetic research. However, much less attention has been paid to the taxa inhabiting the central and eastern Palaearctic, particularly Central Asia, where almost 50% of the species diversity of the genus occurs. Yet, many taxonomic conundrums hinder a proper assessment of the true diversity in these moths. One still unresolved group of species includes Hyles hippophaes and Hyles chamyla. Despite a largely overlapping morphology and ecology, a plethora of infraspecific taxa display some unique divergent characters over a wide geographical area. In this study, we undertook a taxonomic assessment of each population and resolved this species complex using an integrative approach. A combination of new computational techniques (DAISY-II) in comparative morphology and recent advances in DNA extraction methods and sequencing of museum specimens (WISC) alongside more traditional genetic approaches allowed testing of the three main phenotypes—bienerti, chamyla and apocyni—in terms of their morphological, mitochondrial and biogeographical integrity, and to elucidate their evolutionary relationships. Our results support the existence of two closely related species, Hyles chamyla and H. hippophaes, but the former species H. apocyni (here discussed as the ecological form apocyni of H. chamyla) is best regarded as a hybrid between H. chamyla and H. h. bienerti. The results indicate that the evolutionary relationship between H. chamyla and H. hippophaes is one of admixture in the context of ongoing ecological differentiation, which has led to shared morphological characters and a blurring of the species boundaries. These results clarify the evolutionary relationships of this species complex and open future research lines, including the analysis of nuclear markers and denser sampling, particularly of H. hippophaes and H. vespertilio in western Europe.

Microcins are low-molecular-weight, ribosomally produced, highly stable, bacterial-inhibitory molecules involved in competitive, and amensalistic interactions between Enterobacteriaceae in the intestine. These interactions take place in a highly complex chemical landscape, the intestinal eco-active chemosphere, composed of chemical substances that positively or negatively influence bacterial growth, including those originated from nutrient uptake, and those produced by the action of the human or animal host and the intestinal microbiome. The contribution of bacteria results from their effect on the host generated molecules, on food and digested food, and organic substances from microbial origin, including from bacterial degradation. Here, we comprehensively review the main chemical substances present in the human intestinal chemosphere, particularly of those having inhibitory effects on microorganisms. With this background, and focusing on Enterobacteriaceae , the most relevant human pathogens from the intestinal microbiota, the microcin's history and classification, mechanisms of action, and mechanisms involved in microcin's immunity (in microcin producers) and resistance (non-producers) are reviewed. Products from the chemosphere likely modulate the ecological effects of microcin activity. Several cross-resistance mechanisms are shared by microcins, colicins, bacteriophages, and some conventional antibiotics, which are expected to produce cross-effects. Double-microcin-producing strains (such as microcins MccM and MccH47) have been successfully used for decades in the control of pathogenic gut organisms. Microcins are associated with successful gut colonization, facilitating translocation and invasion, leading to bacteremia, and urinary tract infections. In fact, Escherichia coli strains from the more invasive phylogroups (e.g., B2) are frequently microcinogenic. A publicly accessible APD3 database http://aps.unmc.edu/AP/ shows particular genes encoding microcins in 34.1% of E. coli strains (mostly MccV, MccM, MccH47, and MccI47), and much less in Shigella and Salmonella (<2%). Some 4.65% of Klebsiella pneumoniae are microcinogenic (mostly with MccE492), and even less in Enterobacter or Citrobacter (mostly MccS). The high frequency and variety of microcins in some Enterobacteriaceae indicate key ecological functions, a notion supported by their dominance in the intestinal microbiota of biosynthetic gene clusters involved in the synthesis of post-translationally modified peptide microcins. [ABSTRACT FROM AUTHOR]

We present SymPortal (SymPortal.org), a novel analytical framework and platform for genetically resolving the algal symbionts of reef corals using next‐generation sequencing (NGS) data of the ITS2 rDNA. Although the ITS2 marker is widely used to genetically characterize taxa within the family Symbiodiniaceae (formerly the genus Symbiodinium), the multicopy nature of the marker complicates its use. Commonly, the intragenomic diversity resultant from this multicopy nature is collapsed by analytical approaches, thereby focusing on only the most abundant sequences. In contrast, SymPortal employs logic to identify within‐sample informative intragenomic sequences, which we have termed 'defining intragenomic variants' (DIVs), to identify ITS2‐type profiles representative of putative Symbiodiniaceae taxa. By making use of this intragenomic ITS2 diversity, SymPortal is able to resolve genetic delineations using the ITS2 marker at a level that was previously only possible by using additional genetic markers. We demonstrate this by comparing this novel approach to the most commonly used alternative approach for NGS ITS2 data, the 97% similarity clustering to operational taxonomic units (OTUs). The SymPortal platform accepts NGS raw sequencing data as input to provide an easy‐to‐use, standardization‐enforced, and community‐driven framework that integrates with a database to gain resolving power with increased use. We consider that SymPortal, in conjunction with ongoing large‐scale sampling and sequencing efforts, should play an instrumental role in making future sampling efforts more comparable and in maximizing their efficacy in working towards the classification of the global Symbiodiniaceae diversity. [ABSTRACT FROM AUTHOR]

Recent carnivore distributions in eastern Pakistan are poorly documented. In an effort to fill knowledge gaps, we combined field and laboratory molecular approaches to identify the large- and mid-sized mammalian carnivores in Murree, Kotli Sattian, and Kahuta National Park in Rawalpindi, Punjab. A total of 23 sites, each with a transect of 1–4 km in length, were identified and visited monthly over 24 months in 2016–2018. Fecal samples (n = 317) were collected and identified to species in the field based on morphology, odor, and additional signs. A subset (n = 150) of these samples was genetically identified to species using primers designed to amplify an informative region of the canid, felid, mustelid, and viverrid mitochondrial control region. Identified carnivore species for the park include leopard, leopard cat, jungle cat, red fox, golden jackal, small Indian civet, and Canis spp. For leopard, red fox, and golden jackal, field identification rates were accurate at a true-positive rate ≥ 63%. However, there was a notably high rate of field misidentification of putative jungle cat and yellow-throated marten samples, most of which were genetically identified as leopard cat or non-Carnivora samples. We mapped fecal sample presence points based on both morphological and genetically confirmed samples. For five of six dyadic combinations of pairwise species segregation patterns, there was evidence of high rates of non-random co-occurrence of red fox and jackal (p ≤ 0.05). For the other five pairwise combinations, no statistical support was found for non-random distribution. Such findings reiterate the need to combine field and molecular approaches when using fecal samples to survey communities, and underscore the need for broader studies of the carnivore communities of Pakistan. [ABSTRACT FROM AUTHOR]

Chironomus riparius is a well‐established model organism in various fields such as ecotoxicology and ecology, and therefore, environmental preferences, ecological interactions and metabolic traits are well‐studied. With the recent publication of a high‐quality draft genome, as well as different population genetic parameters such as mutation and recombination rate, the species can be used as an alternative to the Drosophila models in experimental population genomics or molecular ecology. To facilitate access to this promising experimental model species for a wider range of researchers, we describe experimental methods to first create and sustain long‐term cultures of C. riparius and then use them to perform repeatable and comparable experiments for various research questions. [ABSTRACT FROM AUTHOR]

The occurrence of plant-associated oomycetes in natural ecosystems and particularly during long-term ecosystem development is largely unknown. Using DNA sequencing, we investigated the frequency and host relationships of plant-root-associated oomycete communities along a 120 000 y glacial chronosequence, comprising site ages with rapid compositional change ("early succession"; 5–70 y old soil); relatively stable higher-diversity sites ("mature", 280–12000 y); and ancient, nutrient-limited soils with declining plant biomass and stature ("retrogression", 60 000, 120 000 y). Plant-associated oomycetes were frequent in early successional sites, occurring in 38–65% of plant roots, but rare (mean 3%) in all older ecosystems. Oomycete OTUs occurred non-randomly with plant host species, and were more frequent on those plant species that declined most strongly in abundance between ecosystem ages. While oomycetes were common in early succession, their absence in older sites suggests a limited role in later stages of ecosystem development. [ABSTRACT FROM AUTHOR]

Genomic technologies have advanced rapidly in recent decades and have ushered in an exciting new era for the study of mammals. Mammalogists working with non-model species are now able to explore many new research areas that were unimaginable only a short time ago. For example, it is currently possible for individual researchers to sequence and assemble genomes for mammalian species of their choosing and to generate genome-wide DNA sequence data from hundreds of individuals simultaneously. Moreover, the miniaturization of both PCR and sequencing technology has created new opportunities for generating real-time DNA or RNA sequence data in the field. Here, we highlight emerging genomic technologies and associated methods that are providing exciting new research avenues for mammalogists in the fields of evolutionary genomics and ecology. Our aim is to not only provide a resource for novel experimental design, but also to provide a record of the current state of genomic applications as the American Society of Mammalogists enters a new century of discovery. [ABSTRACT FROM AUTHOR]

Polyploids are organisms whose genomes consist of more than two complete sets of chromosomes. Both autopolyploids and allopolyploids may display polysomic inheritance. A peculiarity of polysomic inheritance is multivalent formation during meiosis resulting in double-reduction, which occurs when sister chromatid fragments segregate into the same gamete. Double-reduction can result in gametes carrying identical-by-descent alleles and slightly increasing homozygosity. This will cause the genotypic frequencies to deviate from expected values and will thus bias the results of standard population genetic analytical methods used in molecular ecology and selective breeding. In this study, we extend existing double-reduction models to account for any even level of ploidy, and derive the symbolic expressions for genotypic frequencies via two methods. Inbreeding coefficients and heterozygosity under doublereduction and inbreeding are also calculated. Numerical solutions obtained by computer simulations are compared with analytical solutions predicted by the model to validate the model. [ABSTRACT FROM AUTHOR]

Genetic samples can be used to understand and predict the behaviour of species living in a fragmented and temporally changing environment. In this regard, models of coalescence conditioned to an environment through an explicit modelling of population growth and migration have been developed in recent years, and simulators implementing these models have been developed, enabling biologists to estimate parameters of interest with Approximate Bayesian Computation techniques. However, model choice remains limited, and developing new coalescence simulators is extremely time consuming because code re‐use is limited. We present Quetzal, a C++ library composed of re‐usable components, which is sufficiently general to efficiently implement a wide range of spatially explicit coalescence‐based environmental models of population genetics and to embed the simulation in an Approximate Bayesian Computation framework. Quetzal is not a simulation program, but a toolbox for programming simulators aimed at the community of scientific coders and research software engineers in molecular ecology and phylogeography. This new code resource is open‐source and available at https://becheler.github.io/pages/quetzal.html along with other documentation resources. [ABSTRACT FROM AUTHOR]

Nassellaria are marine protists belonging to the Radiolaria lineage (Rhizaria). Their skeleton, made of opaline silica, exhibit an excellent fossil record, extremely valuable in micro-paleontological studies for paleo-environmental reconstruction. Yet, to date very little is known about the extant diversity and ecology of Nassellaria in contemporary oceans, and most of it is inferred from their fossil record. Here we present an integrative classification of Nassellaria based on taxonomical marker genes (18S and 28S ribosomal DNA) and morphological characteristics obtained by optical and scanning electron microscopy imaging. Our phylogenetic analyses distinguished 11 main morpho-molecular clades relying essentially on the overall morphology of the skeleton and not on internal structures as previously considered. Using fossil calibrated molecular clock we estimated the origin of Nassellaria among radiolarians primitive forms in the Devonian (ca. 420 Ma), that gave rise to living nassellarian groups in the Triassic (ca. 250 Ma), during the biggest diversification event over their evolutionary history. This morpho-molecular framework provides both a new morphological classification easier to identify under light microscopy and the basis for future molecular ecology surveys. Altogether, it brings a new standpoint to improve our scarce understanding of the ecology and worldwide distribution of extant nassellarians. [ABSTRACT FROM AUTHOR]

Disentangling pathways by which nonindigenous species expand and spread regionally remains challenging. Molecular ecology tools are often employed to determine the origins and spread of introduced species, but the complexities of some organisms may be reducing the efficacy of these tools. Some colonial species exhibit complexities by way of chimerism and winter colony regression, which may alter the genetic diversity of populations and mask the connectivity occurring among them. This study uses nuclear microsatellite data and simple GIS‐based modeling to investigate the influence of chimerism and winter regression on the genetic diversity and patterns of genetic population connectivity among colonies of Didemnum vexillum on artificial substrates. Colonies sampled in summer were shown to form a metapopulation, with high levels of admixture, extreme outcrossing, and some substructure. These patterns were consistent within the subsampled winter colonies and with the inclusion of chimeric data. However, allelic richness and diversity were significantly different between winter and summer samples, altering interpretations relating to population connectivity and pelagic larval duration. This study demonstrates the importance of including seasonal sampling and imperative life history traits in genetic studies for clear interpretations and the successful management of introduced species. [ABSTRACT FROM AUTHOR]

Poor conditions during early development can initiate trade‐offs that favour current survival at the expense of somatic maintenance and subsequently, future reproduction. However, the mechanisms that link early and late life‐history are largely unknown. Recently it has been suggested that telomeres, the nucleoprotein structures at the terminal end of chromosomes, could link early‐life conditions to lifespan and fitness. In wild purple‐crowned fairy‐wrens, we combined measurements of nestling telomere length (TL) with detailed life‐history data to investigate whether early‐life TL predicts fitness prospects. Our study differs from previous studies in the completeness of our fitness estimates in a highly philopatric population. The association between TL and survival was age‐dependent with early‐life TL having a positive effect on lifespan only among individuals that survived their first year. Early‐life TL was not associated with the probability or age of gaining a breeding position. Interestingly, early‐life TL was positively related to breeding duration, contribution to population growth and lifetime reproductive success because of their association with lifespan. Thus, early‐life TL, which reflects growth, accumulated early‐life stress and inherited TL, predicted fitness in birds that reached adulthood but not noticeably among fledglings. These findings suggest that a lack of investment in somatic maintenance during development particularly affects late life performance. This study demonstrates that factors in early‐life are related to fitness prospects through lifespan, and suggests that the study of telomeres may provide insight into the underlying physiological mechanisms linking early‐ and late‐life performance and trade‐offs across a lifetime. [ABSTRACT FROM AUTHOR]

Abstract Wastewater treatment plants (WWTPs) are the main hotspots for the release of antibiotics, including the widely used sulphonamides. Microbes play important roles in eliminating sulphonamides in WWTPs, and knowledge about these degraders and their interactions within the microbial community is crucial for operating and optimising WWTPs. In the present study, stable isotope probing (SIP) coupled with high-throughput sequencing as culture-independent approach revealed four operational taxonomic units (OTUs) involved in sulphamethoxazole (SMX) degradation in activated sludge. Except for the OTU affiliated with Gammaproteobacteria, the others have not been previously reported to possess the ability to metabolise SMX. The isolated SMX degrader by culture-dependent method did not participate in SMX biodegradation in situ according to the SIP analysis, and showed weak correlations with other members in the activated sludge. The complex interactions between in situ active SMX degraders and non-degrading microbes might explain our failure to isolate these degraders. In addition, sul1 genes associated with SMX resistance were also labelled with 13C, suggesting that they might benefit from SMX degradation and/or originate from the active SMX degraders. These findings broaden our understanding of the diversity of SMX-degrading microbes and their associated characteristics in WWTPs. Graphical abstract Unlabelled Image Highlights • SIP coupled with MENA was firstly applied to study the SMX degraders in situ. • Planctomycetaceae , GP7 and incertae sedis were firstly linked with SMX degradation. • Active SMX-degraders hold intense connections with various bacteria in the community. • The sul1 gene might be hosted by the active SMX-degraders. [ABSTRACT FROM AUTHOR]

Background: High-throughput amplicon sequencing of environmental DNA (eDNA metabarcoding) has become a routine tool for biodiversity survey and ecological studies. By including sample-specific tags in the primers prior PCR amplification, it is possible to multiplex hundreds of samples in a single sequencing run. The analysis of millions of sequences spread into hundreds to thousands of samples prompts for efficient, automated yet flexible analysis pipelines. Various algorithms and software have been developed to perform one or multiple processing steps, such as paired-end reads assembly, chimera filtering, Operational Taxonomic Unit (OTU) clustering and taxonomic assignment. Some of these software are now well established and widely used by scientists as part of their workflow. Wrappers that are capable to process metabarcoding data from raw sequencing data to annotated OTU-to-sample matrix were also developed to facilitate the analysis for non-specialist users. Yet, most of them require basic bioinformatic or command-line knowledge, which can limit the accessibility to such integrative toolkits. Furthermore, for flexibility reasons, these tools have adopted a step-by-step approach, which can prevent an easy automation of the workflow, and hence hamper the analysis reproducibility. Results: We introduce SLIM, an open-source web application that simplifies the creation and execution of metabarcoding data processing pipelines through an intuitive Graphic User Interface (GUI). The GUI interact with well-established software and their associated parameters, so that the processing steps are performed seamlessly from the raw sequencing data to an annotated OTU-to-sample matrix. Thanks to a module-centered organization, SLIM can be used for a wide range of metabarcoding cases, and can also be extended by developers for custom needs or for the integration of new software. The pipeline configuration (i.e. the modules chaining and all their parameters) is stored in a file that can be used for reproducing the same analysis. Conclusion: This web application has been designed to be user-friendly for non-specialists yet flexible with advanced settings and extensibility for advanced users and bioinformaticians. The source code along with full documentation is available on the GitHub repository (https://github.com/yoann-dufresne/SLIM) and a demonstration server is accessible through the application website (https://trtcrd.github.io/SLIM/). [ABSTRACT FROM AUTHOR]

Parentage analysis is a cornerstone of molecular ecology that has delivered fundamental insights into behaviour, ecology and evolution. Microsatellite markers have long been the king of parentage, their hypervariable nature conferring sufficient power to correctly assign offspring to parents. However, microsatellite markers have seen a sharp decline in use with the rise of next‐generation sequencing technologies, especially in the study of population genetics and local adaptation. The time is ripe to review the current state of parentage analysis and see how it stands to be affected by the emergence of next‐generation sequencing approaches. We find that single nucleotide polymorphisms (SNPs), the typical next‐generation sequencing marker, remain underutilized in parentage analysis but are gaining momentum, with 58 SNP‐based parentage analyses published thus far. Many of these papers, particularly the earlier ones, compare the power of SNPs and microsatellites in a parentage context. In virtually every case, SNPs are at least as powerful as microsatellite markers. As few as 100–500 SNPs are sufficient to resolve parentage completely in most situations. We also provide an overview of the analytical programs that are commonly used and compatible with SNP data. As the next‐generation parentage enterprise grows, a reliance on likelihood and Bayesian approaches, as opposed to strict exclusion, will become increasingly important. We discuss some of the caveats surrounding the use of next‐generation sequencing data for parentage analysis and conclude that the future is bright for this important realm of molecular ecology. [ABSTRACT FROM AUTHOR]

The current state of knowledge of the suprageneric relationships in Cholevinae is either derived from informal evaluations of putative synapomorphies or based on molecular studies with limited taxonomic sampling. Here we assessed the higher‐level relationships in this subfamily based on a phylogenetic analysis of 97 morphological characters scored for 93 terminals, representing all tribes. Both parsimony and Bayesian analyses were used. The monophyletic origin of Cholevinae was corroborated, except for the unexpected inclusion of Leptinus in the implied weighting analysis. Eucatopini + Oritocatopini were retrieved as basal branches in the evolution of Cholevinae. The monophyletic origin of all remaining Cholevinae was confirmed, which is consistent with molecular evidence. Anemadini was non‐monophyletic, in accordance with earlier hypotheses. Cholevini was rendered non‐monophyletic by the uncertain inclusion of Prionochaeta and the consistent exclusion of Cholevinus. A close affinity of Ptomaphagini to Sciaphyini and Leptodirini was suggested, although the position of Sciaphyes remains uncertain. The phylogenetic hypothesis of Cholevinae provided here is the most comprehensive presently available. The list of characters shows that a substantial part of the data was obtained from the ventral side. This is a strong argument for a detailed pictorial documentation of the ventral body parts in taxonomic descriptions, in contrast to the common practice of only illustrating the dorsal habitus of the beetles. [ABSTRACT FROM AUTHOR]

Highlights • Biochar addition increased thermophilic temperature and extended the high temperature period of compost. • Biochar addition enriched more amounts of specific bacterial taxa related to decomposition of organic matter. • The microbial co-occurrence pattern and keystone taxa were observed during composting. • DOC and TN were the key environmental factors for community succession. Abstract High-throughput sequencing and network analysis were used to investigate the dynamic changes and the effects on bacterial community structures in green waste compost after biochar addition. Biochar addition led to higher thermophilic temperatures and total nitrogen (TN) concentrations and a longer thermophilic period compared with no biochar addition. Biochar also greatly influenced the composition of the bacterial community. Nitriliruptoraceae and Bacillaceae abundances increased in the poultry manure after biochar treatment, and Alcaligenaceae , Rhodispirillaceae and Xanthomonadaceae were more abundant in the cattle manure. Dissolved organic carbon (DOC) and TN emerged as the key determinants of bacterial community composition. Network analysis revealed that DOC had strong positive associations with some taxa (e.g. Comamonas , Leucobacter and Acidimicrobiales), whereas TN had negative associations with other taxa (e.g. Microbacteriaceae and Aeromicrobium). This study has revealed the key taxa related to the carbon and nitrogen cycle during composting with biochar. [ABSTRACT FROM AUTHOR]

The article discusses developments in the molecular ecology-focused journal. Topics discussed include the periodical's top ranking among publications in the field of evolution and ecology, the recipients of the periodical's Molecular Ecology Prize and Harry Smith Prize, and the decision of professor Louis Bernatchez to step down as the reviews editor. Also discussed are changes in the commissioning of special issues and the performance of the Molecular Ecologist blog.

The article presents a profile of scientist and geneticist Robin S. Waples, the recipient of the 2018 Molecular Ecology Prize. Topics discussed include Waples' career as a senior scientist at the Northwest Fisheries Science Center (NWFSC) of the U.S. National Oceanic and Atmospheric Administration's (NOAA) National Marine Fisheries Service, his research on conservation genetics and population genetics of marine species, and other awards he received including the NOAA Administrator's Award.

Abstract Understanding the role of microbes in the solubility of cadmium (Cd) is of fundamental importance for remediation of Cd toxicity. The present study aimed to identify the microbes that involved in regulating Cd solubility and to reveal possible mechanisms. Therefore, microbial communities were investigated through high-throughput sequencing approach, the molecular ecological network was constructed and metagenomes were predicted. Our results indicated that redox conditions affected both the solubility of soil Cd and the microbial communities. Anaerobic microbes, such as Anaerolineaceae, did not only play important roles in shaping the microbial community in soils, but might also be involved in regulating the Cd solubility. Two possible mechanisms that how Anaerolineaceae involved in Cd solubility are (1) Anaerolineaceae are important organic matter degraders under anoxic conditions and (2) Anaerolineaceae can co-exist with methane metabolism microbes, while methane metabolism promotes the precipitation of soluble Cd. Thus, application of Anaerolineaceae in bioremediation of soil Cadmium contamination is a potential approach. The study provided a novel insight into the role of microbial community in the regulation of Cd solubility under different redox conditions, and suggested a potential approach for the remediation of soil Cd contamination. Graphical abstract Unlabelled Image [ABSTRACT FROM AUTHOR]

Abstract The application of next generation sequencing and molecular ecology to the systematics and taxonomy of prokaryotes offers enormous insights into prokaryotic biology. This discussion explores some major disagreements but also considers the opportunities associated with the nomenclature of the uncultured taxa, the use of genome sequences as type material, the plurality of the nomenclatural code, and the roles of an official or computer-assisted taxonomy. [ABSTRACT FROM AUTHOR]

Development of effective metabarcoding-based tools for ecological assessment requires more than just detailed knowledge of ecology and molecular genetics. There is also a need to understand the context within which they will be used, and for the organisation that uses it to understand the techniques involved and, more especially, how the data that are produced differs from that generated by traditional ecological methods. Lessons learnt during the development of a metabarcoding tool for phytobenthos in the UK are set out in this paper. This attempted to develop a molecular "mirror" of the existing light microscopy-based approach to ecological assessment. Although this conservative approach does not exploit the full potential of metabarcoding data, it does mean that benchmarks exist against which performance and data can be judged. However, the pace of developments within molecular ecology means that regulators will need to find ways of incorporating new scientific insights whilst, at the same time, ensuring a stable regulatory regime. Installation of a metabarcoding technique within a regulatory organisation, in other words, is more than a transaction in which one approach is switched for another. A deeper transformation of the organisation is required. [ABSTRACT FROM AUTHOR]

In the context of antimicrobial resistance as one of the most serious issues faced globally by health providers, we explored a practical introduction to molecular microbial ecology. We designed field work and practical experiments for third year members of a 4 year undergraduate Masters Program, in which the students employed traditional and novel isolation techniques to identify antimicrobial activities from soil dwelling microorganisms. Students gained experience in isolating DNA from complex microbial communities, amplifying 16S rRNA genes and applied richness/diversity indices as well as principal coordinate analyses to the interpretation of the data they obtained from high throughput sequencing. Our results confirmed that isolation chips facilitate the growth of a greater diversity and different species subset from the complex soil microorganism community than traditional plate spreading techniques. However, rarefaction of 16S rRNA amplicon sequencing data showed that the majority of observed species in soil remain unculturable by current methods. Based on the written reports produced by the students carrying out the work, we concluded that the described protocols are robust and informative, that these activities provide a good practical introduction to the theories and practice of molecular ecology and can be easily deployed to groups of six or more students in a cost‐effective manner. We explored a practical introduction to molecular microbial ecology through field work and laboratory experiments for third‐year students in a 4‐year undergraduate Masters program. Students isolated DNA from complex microbial communities and applied richness / diversity indices as well as principal coordinates analyses to interpret the 16S rRNA data they obtained from high‐throughput sequencing. These activities provided a good practical introduction to the theories and practice of molecular ecology and were easy to deploy in a cost‐effective manner to groups of six or more students. [ABSTRACT FROM AUTHOR]

Fish can adapt to a wide range of temperature changes, however, the molecular mechanisms that underlie these adaptations are not fully understood. In this study, fluorescently labeled differential display (FDD) technique was used to detect mRNAs that are differentially expressed in hepatic tissues of Tilapia zillii fish subjected to a temperature reduction regime that combined both accelerated and stationary cold stresses. The two arbitrary primers (DD1 and DD2) with the Oligo d(T)23-C generated a total of 18 and 37 unknown messenger RNA (UMR) bands, respectively. Both DD1 & DD2 arbitrary primers with oligo d(T)23-C scored four stress-related unidentified fragments after refining. Using the two arbitrary primers with the Oligo d(T)23-G, a total of 371 and 372 UmRs were yielded, respectively. Both arbitrary primers scored nine stress-related unidentified fragments after refining. All UmR were tested for its correlation to time and temperature factors scaled for the current experiment, only four were found to be correlated for both factors. Thus, T. zillii responded to temperature reduction by adjusting the expression of a set number of genes that may be required for their adaptation and tolerance to low environmental temperature.