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At least 415 records · Page 23Linked to original sources

Quantitative PCR detection of endangered diamond darter Crystallaria Cincotta in environmental DNA: Employing locked nucleic acids and blocking probe for specificity

This study presents a quantitative PCR (qPCR) assay for the detection of the endangered diamond darter Crystallaria cincotta from environmental DNA (eDNA) in water samples. The assay design is based on an alignment of mitochondrial cytochrome b DNA sequences from 58 individuals representing 25 percid species. Leveraging genetic differences, a species-specific qPCR assay was designed, incorporating alocked nucleic acid (LNA)-enriched probe and a secondary blocking probe to enhance specificity. The assay targets a 93-base pair fragment that includes a diagnostic single nucleotide polymorphism in the probe region; combined with multiple primer mismatches, this provides specificity for distinguishing C. cincotta from other sympatric percid species. Specificity was validated by testing genomic DNA from 16 percid species and synthetic templates, confirming no cross-reactivity. Performance metrics, including the standard curve, qPCR efficiency, limit of detection, and limit of quantification, are reported. The qPCR assay exhibited sufficient sensitivity to detect C. cincotta eDNA in environmental water samples collected from occupied riverine habitats. This study illustrates the effectiveness of LNA-enriched and blocking probes in developing species-specific qPCR assays for eDNA applications, demonstrating their utility in accurately distinguishing closely related species within diverse fish communities.

Conservation Genetics Resources↗

A critical review of bioaccumulation and biotransformation of organic chemicals in birds

A literature review of bioaccumulation and biotransformation of organic chemicals in birds was undertaken, aiming to support scoping and prioritization of future research. The objectives were to characterize available bioaccumulation/biotransformation data, identify knowledge gaps, determine how extant data can be used, and explore the strategy and steps forward. An intermediate approach balanced between expediency and rigor was taken given the vastness of the literature. Following a critical review of > 500 peer-reviewed studies, > 25,000 data entries and 2 million information bytes were compiled on > 700 organic compounds for ~ 320 wild species and 60 domestic breeds of birds. These data were organized into themed databases on bioaccumulation and biotransformation , field survey , microsomal enzyme activity , metabolic pathway , and bird taxonomy and diet . Significant data gaps were identified in all databases at multiple levels. Biotransformation characterization was largely fragmented over metabolite/pathway identification and characterization of enzyme activity or biotransformation kinetics. Limited biotransformation kinetic data constrained development of an avian biotransformation model. A substantial shortage of in vivo biotransformation kinetics has been observed as most reported rate constants were derived in vitro. No metric comprehensively captured all key contaminant classes or chemical groups to support broad-scope modeling of bioaccumulation or biotransformation. However, metrics such as biota-feed accumulation factor, maximum transfer factor, and total elimination rate constant were more readily usable for modeling or benchmarking than other reviewed parameters. Analysis demonstrated the lack of bioaccumulation/biotransformation characterization of shorebirds, seabirds, and raptors. In the study of bioaccumulation and biotransformation of organic chemicals in birds, this review revealed the need for greater chemical and avian species diversity, chemical measurements in environmental media, basic biometrics and exposure conditions, multiple tissues/matrices sampling, and further exploration on biotransformation. Limitations of classical bioaccumulation metrics and current research strategies used in bird studies were also discussed. Forward-looking research strategies were proposed: adopting a chemical roadmap for future investigations, integrating existing biomonitoring data, gap-filling with non-testing approaches, improving data reporting practices, expanding field sampling scopes, bridging existing models and theories, exploring biotransformation via avian genomics, and establishing an online data repository.

Reviews of Environmental Contamination and Toxicol↗

The genetic diversity and epizootiology of infectious hematopoietic necrosis virus

Infectious hematopoietic necrosis virus (IHNV) is a rhabdovirus which causes a serious disease in salmondd fish. The T1 ribonuclease fingerprinttin method was used to compare the RNA genomes of 26 isolates of IHNV recovered from sockeye salmon ( Oncorhynchus nerka ), chinook salmon ( O. tshawytscha ), and steelhead trout ( O. mykiss ) throughout the enzootic portion of western North America. Most of the isolates as a source of genetic variation. In from a single year (1987) to limit time of isolation as a source of genetic variation. In addition, isolates from different years collected at three sites were analyzed to investigate genetic drift or evolution of IHNV within specific locations. All of the isolates examined by T1 fingerprint analysis contained less than a 50% variation in spot location and were represented by a single fingerprint group. The observed variation was estimated to correspond to less than 5% variation in the nucleic acid sequence. However, sufficient variation was detected to separate the isolates into four subgroups which appeared to correlate to different geographic regions. Host species appeared not to be a significant source of variation. The evolutionary and epizootiologic significance of these findings and their relationship to other evidence of genetic variation in IHNV isolates are discussed.

Virus Research↗

The glycoprotein genes and gene junctions of the fish rhabdoviruses spring viremia of carp virus and hirame rhabdovirus: Analysis of relationships with other rhabdoviruses

The nucleotide sequences of the glycoprotein genes and all of the internal gene junctions of the fish pathogenic rhabdoviruses spring viremia of carp virus (SVCV) and hirame rhabdovirus (HIRRV) have been determined from cDNA clones generated from viral genomic RNA. The SVCV glycoprotein gene sequence is 1588 nucleotides (nt) long and encodes a 509 amino acid (aa) protein. The HIRRV glycoprotein gene sequence comprises 1612 nt, coding for a 508 aa protein. In sequence comparisons of 15 rhabdovirus glycoproteins, the SVCV glycoprotein gene showed the highest amino acid sequence identity (31.2-33.2%) with vesicular stomatitis New Jersey virus (VSNJV), Chandipura virus (CHPV) and vesicular stomatitis Indiana virus (VSIV). The HIRRV glycoprotein gene showed a very high amino acid sequence identity (74.3%) with the glycoprotein gene of another fish pathogenic rhabdovirus, infectious hematopoietic necrosis virus (IHNV), but no significant similarity with glycoproteins of VSIV or rabies virus (RABV). In phylogenetic analyses SVCV was grouped consistently with VSIV, VSNJV and CHPV in the Vesiculovirus genus of Rhabdoviridae. The fish rhabdoviruses HIRRV, IHNV and viral hemorrhagic septicemia virus (VHSV) showed close relationships with each other, but only very distant relationships with mammalian rhabdoviruses. The gene junctions are highly conserved between SVCV and VSIV, well conserved between IHNV and HIRRV, but not conserved between HIRRV/IHNV and RABV. Based on the combined results we suggest that the fish lyssa-type rhabdoviruses HIRRV, IHNV and VHSV may be grouped in their own genus within the family Rhabdoviridae. Aquarhabdovirus has been proposed for the name of this new genus.

Virus Research↗

Reassociation and hybridization properties of DNAs from several species of fish

Reassociation and hybridization properties from spectrophotometric studies of DNAs from 10 species of fish indicate: 1. Great diversity in the amounts of repeated sequences in the genomes of different species - more specialized fish had less redundancy. 2. Large differences in the complexities of the DNAs - more specialized fish had less information. 3. Little homology between sequences of remotely related species but substantial homology between sequences of closely related species.

Comparative Biochemistry and Physiology, Part B: C↗

Hormonal control of euryhalinity

Hormones play a critical role in maintaining body fluid balance in euryhaline fishes during changes in environmental salinity. The neuroendocrine axis senses osmotic and ionic changes, then signals and coordinates tissue-specific responses to regulate water and ion fluxes. Rapid-acting hormones, e.g. angiotensins, cope with immediate challenges by controlling drinking rate and the activity of ion transporters in the gill, gut, and kidney. Slow-acting hormones, e.g. prolactin and growth hormone/insulin-like growth factor-1, reorganize the body for long-term acclimation by altering the abundance of ion transporters and through cell proliferation and differentiation of ionocytes and other osmoregulatory cells. Euryhaline species exist in all groups of fish, including cyclostomes, and cartilaginous and teleost fishes. The diverse strategies for responding to changes in salinity have led to differential regulation and tissue-specific effects of hormones. Combining traditional physiological approaches with genomic, transcriptomic, and proteomic analyses will elucidate the patterns and diversity of the endocrine control of euryhalinity.

Book chapter↗

Smolt physiology and endocrinology

Hormones play a critical role in maintaining body fluid balance in euryhaline fishes during changes in environmental salinity. The neuroendocrine axis senses osmotic and ionic changes, then signals and coordinates tissue-specific responses to regulate water and ion fluxes. Rapid-acting hormones, e.g. angiotensins, cope with immediate challenges by controlling drinking rate and the activity of ion transporters in the gill, gut, and kidney. Slow-acting hormones, e.g. prolactin and growth hormone/insulin-like growth factor-1, reorganize the body for long-term acclimation by altering the abundance of ion transporters and through cell proliferation and differentiation of ionocytes and other osmoregulatory cells. Euryhaline species exist in all groups of fish, including cyclostomes, and cartilaginous and teleost fishes. The diverse strategies for responding to changes in salinity have led to differential regulation and tissue-specific effects of hormones. Combining traditional physiological approaches with genomic, transcriptomic, and proteomic analyses will elucidate the patterns and diversity of the endocrine control of euryhalinity.

Book chapter↗

Hepeviruses of aquatic organisms

Originally reported in California, the cutthroat trout virus (CTV) has now been isolated from nine species of salmonids in North America. Early work focused on the replication and physical characteristics of the virus, but 20 years later was determined to be most closely related to the hepatitis E virus. The small genome is positive-sense, single-stranded RNA similar to other members of the family Hepeviridae , which now contains its own genus Piscihepevirus with two distinct genotypes, CTV-1 and CTV-2. While CTV has not been associated with acute disease in fish, the virus could form persistently infected cell cultures that may aid research in treating hepatitis E-like viruses affecting humans or other animals. Interestingly, trout exposed to CTV were protected for about a month against subsequent exposure to the infectious hematopoietic necrosis virus. Replicating agents suspected to be CTV can be confirmed by polymerase chain reaction (PCR), quantitative PCR, and sequencing. Other unclassified hepeviruses detected in fish using viral metagenomics include Wenling fish hepevirus, Wenling moray eel hepevirus, Murray–Darling carp hepevirus, and eastern mosquitofish hepevirus. The family Hepeviridae has been placed in the order Hepevirales together with the family Matonaviridae (rubella virus), with member viruses having amino acid homology in the helicase and replicase regions of the nonstructural proteins. In addition, using next-generation sequencing, a hepe-like sequence was characterized in diseased giant freshwater prawn Macrobrachium rosenbergii and named Crustacea hepe-like virus 1. Thus the family Hepeviridae continues to expand among aquatic animal hosts.

Book chapter↗

Using quantitative polymerase chain reaction to assess phytoplankton and indicate eutrophication in freshwater rivers: A multiyear nationwide study across the United States

Phytoplankton are essential primary producers in fresh surface water that are critical to the health of ecosystems. However, phytoplankton overgrowth due to eutrophication threatens ecological, economic, and public health. Therefore, assessing phytoplankton is fundamental for understanding the productivity, health, and trophic status of freshwater ecosystems. Light microscopy and chlorophyll a assessment are common approaches for studying phytoplankton. They are easy to use, cost-effective, and reliable but have significant limitations. Microscopy has a low throughput and is time-consuming and labor-intensive. Chlorophyll a assessment does not reveal phytoplankton community composition and structure. For comparison, quantitative polymerase chain reaction (qPCR) is widely applied in quantifying microorganisms, offering multiple advantages, including high throughput, sensitivity, accuracy, and robustness. However, a research gap remains regarding the feasibility of using qPCR to assess phytoplankton and indicate trophic status of freshwater bodies. We conducted a nationwide, multiyear study in the United States to compare the performance of qPCR, microscopy, and chlorophyll a assessment in assessing phytoplankton and trophic statuses of multiple freshwater rivers. From early summer to late fall in 2017, 2018, and 2019, we assessed phytoplankton, chlorophyll a , pheophytin a , and the overall Trophic Level Index ( TLI Overall ) at the sampling sites in 12 large freshwater rivers in three regions (western, midcontinent, and eastern) across the United States. The seasonal summed abundance of four major phytoplankton taxa [Bacillariophyta (diatoms), Cyanobacteria (blue-green algae), Chlorophyta (green algae), and Dinoflagellates (Dinophyta)] ranged from 6.88 log 10 (GCN·L –1 ) (the Connecticut River, 2017) to 9.29 log 10 (GCN·L –1 ) (the Kansas River, 2019) (GCN: gene or genome copy number). qPCR- and microscopy-based phytoplankton abundance of eight phytoplankton taxa had a significant positive allometric or log-linear correlation (adjusted R 2 = 0.836, p -value < 0.001, n = 815). In addition, qPCR-based phytoplankton abundance had positive allometric or log-linear correlations with chlorophyll a (adjusted R 2 = 0.5437, p -value < 0.001, n = 164), pheophytin a (adjusted R 2 = 0.3378, p -value < 0.001, n = 164), and TLI Overall (adjusted R 2 = 0.4789, p -value < 0.001, n = 164). Therefore, qPCR is a promising alternative to microscopy and chlorophyll a for studying phytoplankton and trophic status in freshwater rivers. Moreover, phytoplankton abundance had limited temporal variation within each sampling season and over the three sampling seasons in 2017, 2018, and 2019 but showed clear spatial variation. The midcontinent sites had significantly higher phytoplankton abundance, chlorophyll a concentrations, pheophytin a concentrations, and TLI Overall values than those in the eastern and western rivers, reflecting the higher trophic statuses of the midcontinent rivers. This work also provides the thresholds of qPCR-based phytoplankton abundance for delineating trophic statuses in freshwater rivers. Overall, this work demonstrates that qPCR is a promising tool for studying phytoplankton and characterizing the trophic status of freshwater rivers.

Book chapter↗

Genetic analysis of North American Phragmites australis guides management approaches

Phragmites australis subsp. australis is an invasive and ecologically detrimental plant in multiple regions of North America. Its co-occurrence with the native subspecies, and multiple instances of hybridization, has created the need to differentiate Phragmites subspecies or haplotypes so that management can be appropriately targeted to the invader. We compiled a review of current genetic discrimination methods among the three Phragmites subspecies inhabiting the United States and Canada, and discussed how each method can contribute to control of the introduced subspecies while preserving the two endemic subspecies. We also discussed various control tools and the implications of Phragmites genetics for implementation. The Phragmites subspecies endemic to North America have environmental or infrastructure significance (e.g., habitat sustainability, biodiversity, storm surge and erosion protection). Thus, faster and more accurate differentiation among the endemic and introduced subspecies is needed. Additionally, more in-depth genetic information on Phragmites subspecies could support better management decisions, as well as the development of improved control treatments. This review highlights technologies and approaches currently available for genetic identification, recently collected genomic, transcriptomic and proteomic information, and implications for biological control and herbicide treatments.

Aquatic Botany↗

Respiratory arsenate reductase as a bidirectional enzyme

The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe–S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism.

Biochemical and Biophysical Research Communication↗

Emerging and reemerging diseases of avian wildlife

Of the many important avian wildlife diseases, aspergillosis, West Nile virus, avipoxvirus, Wellfleet Bay virus, avian influenza, and inclusion body disease of cranes are covered in this article. Wellfleet Bay virus, first identified in 2010, is considered an emerging disease. Avian influenza and West Nile virus have recently been in the public eye because of their zoonotic potential and links to wildlife. Several diseases labeled as reemerging are included because of recent outbreaks or, more importantly, recent research in areas such as genomics, which shed light on the mechanisms whereby these adaptable, persistent pathogens continue to spread and thrive.

Veterinary Clinics of North America: Exotic Animal↗

Identification, characterization and genetic mapping of TLR7, TLR8a1 and TLR8a2 genes in rainbow trout (Oncorhynchus mykiss)

Induction of the innate immune pathways is critical for early anti-viral defense but there is limited understanding of how teleost fish recognize viral molecules and activate these pathways. In mammals, Toll-like receptors (TLR) 7 and 8 bind single-stranded RNA of viral origin and are activated by synthetic anti-viral imidazoquinoline compounds. Herein, we identify and describe the rainbow trout (Oncorhynchus mykiss) TLR7 and TLR8 gene orthologs and their mRNA expression. Two TLR7/8 loci were identified from a rainbow trout bacterial artificial chromosome (BAC) library using DNA fingerprinting and genetic linkage analyses. Direct sequencing of two representative BACs revealed intact omTLR7 and omTLR8a1 open reading frames (ORFs) located on chromosome 3 and a second locus on chromosome 22 that contains an omTLR8a2 ORF and a putative TLR7 pseudogene. We used the omTLR8a1/2 nomenclature for the two trout TLR8 genes as phylogenetic analysis revealed that they and all the other teleost TLR8 genes sequenced to date are similar to the zebrafish TLR8a, but are distinct from the zebrafish TLR8b. The duplicated trout loci exhibit conserved synteny with other fish genomes extending beyond the tandem of TLR7/8 genes. The trout TLR7 and 8a1/2 genes are composed of a single large exon similar to all other described TLR7/8 genes. The omTLR7 ORF is predicted to encode a 1049 amino acid (aa) protein with 84% similarity to the Fugu TLR7 and a conserved pattern of predicted leucine-rich repeats (LRR). The omTLR8a1 and omTLR8a2 are predicted to encode 1035- and 1034-aa proteins, respectively, and have 86% similarity to each other. omTLR8a1 is likely the ortholog of the only Atlantic salmon TLR8 gene described to date as they have 95% aa sequence similarity. The tissue expression profiles of omTLR7, omTLR8a1 and omTLR8a2 in healthy trout were highest in spleen tissue followed by anterior and then posterior kidney tissues. Rainbow trout anterior kidney leukocytes produced elevated levels of pro-inflammatory and type I interferon cytokines mRNA in response to stimulation with the human TLR7/8 agonist R848 or the TLR3 agonist poly I:C. Only poly I:C-induced IFN2 transcription was significantly suppressed in the presence of chloroquine, a compound known to block endosomal acidification and inhibit endosomal maturation. The effect of chloroquine on R848-induced cytokine expression was equivocal and so it remains questionable whether rainbow trout recognition of R848 requires endosomal maturation. TLR7 and TLR8a1 expression levels in rainbow trout anterior kidney leukocytes were not affected by poly I:C or R848 treatments, but surprisingly, TLR8a2 expression was moderately down-regulated by R848. The down-regulation of omTLR8a2 may imply that this gene has evolved to a new or altered function in rainbow trout, as often occurs when the two duplicated genes remain active.

Developmental and Comparative Immunology↗

Description of an elasmobranch TCR coreceptor: CD8&alpha; from Rhinobatos productus

Cell-mediated immunity plays an essential role for the control and eradication of intracellular pathogens. To learn more about the evolutionary origins of the first signal (Signal 1) for T-cell activation, we cloned CD8α from an elasmobranch, Rhinobatos productus. Similar to full-length CD8α cDNAs from other vertebrates, Rhpr-CD8α (1800 bp) encodes a 219 amino acid open reading frame composed of a signal peptide, an extracellular IgSF V domain and a stalk/hinge region followed by a well-conserved transmembrane domain and cytoplasmic tail. Overall, the mature Rhpr-CD8α protein (201 aa) displays ~30% amino acid identity with mammalian CD8α including absolute conservation of cysteine residues involved in the IgSf V domain fold and dimerization of CD8αα and CD8αβ. One prominent feature is the absence of the LCK association motif (CXC) that is needed for achieving signal 1 in tetrapods. Both elasmobranch and teleost CD8α protein sequences possess a similar but distinctly different motif (CXH) in the cytoplasmic tail. The overall genomic structure of CD8α has been conserved during the course of vertebrate evolution both for the number of exons and phase of splicing. Finally, quantitative RTPCR demonstrated that elasmobranch CD8α is expressed in lymphoid-rich tissues similar to CD8 in other vertebrates. The results from this study indicate the existence of CD8 prior to the emergence of the gnathostomes (>450 MYA) while providing evidence that the canonical LCK association motif in mammals is likely a derived characteristic of tetrapod CD8α, suggesting potential differences for T-cell education and activation in the various gnathostomes.

Developmental and Comparative Immunology↗

The Florida manatee (Trichechus manatus latirostris) immunoglobulin heavy chain suggests the importance of clan III variable segments in repertoire diversity

Manatees are a vulnerable, charismatic sentinel species from the evolutionarily divergent Afrotheria. Manatee health and resistance to infectious disease is of great concern to conservation groups, but little is known about their immune system. To develop manatee-specific tools for monitoring health, we first must have a general knowledge of how the immunoglobulin heavy (IgH) chain locus is organized and transcriptionally expressed. Using the genomic scaffolds of the Florida manatee ( Trichechus manatus latirostris ), we characterized the potential IgH segmental diversity and constant region isotypic diversity and performed the first Afrotherian repertoire analysis. The Florida manatee has low V(D)J combinatorial diversity (3744 potential combinations) and few constant region isotypes. They also lack clan III V segments, which may have caused reduced VH segment numbers. However, we found productive somatic hypermutation concentrated in the complementarity determining regions. In conclusion, manatees have limited IGHV clan and combinatorial diversity. This suggests that clan III V segments are essential for maintaining IgH locus diversity.

Developmental and Comparative Immunology↗

Characterization of immunoglobulin light chain utilization and variable family diversity in rainbow trout

This study characterizes immunoglobulin light chain (IgL) expression and variable family usage in rainbow trout. IgL transcripts were generated by 5’ RACE from both immune and TNP-KLH immunized fish. Phylogenetic analysis revealed that the IgL variable regions clustered into seven different families: three kappa families (two newly described in this study), three sigma families, and a single lambda family. IgL1 and IgL3 transcripts expressing identical variable regions were identified and genomic analysis revealed that the two isotypes are co-localized on chromosomes 7, 15, 18, and 21 allowing for potential rearrangement between clusters. Fish were immunized with TNP-KLH (n = 5) and percent expression of IgL1, IgL2, IgL3, and IgL4 measured by qRT-PCR from immune tissues and magnetically sorted TNP-specific lymphocyte populations. In all samples IgL1 constituted 80–95% of the transcripts. The percentage of anti-TNP specific IgL1 transcripts was measured in naïve, unsorted, and TNP-specific cell populations of TNP-KLH fish (n = 3) and found to be significantly higher in the TNP positive cell population (21%) compared to the naïve population (1%; p = 0.02) suggesting that there is a selection of TNP specific IgL sequences.

Developmental and Comparative Immunology↗

Gene transcription patterns in response to low level petroleum contaminants in Mytilus trossulus from field sites and harbors in southcentral Alaska

The 1989 Exxon Valdez oil spill damaged a wide range of natural resources, including intertidal communities, and post-spill studies demonstrated acute and chronic exposure and injury to an array of species. Standard toxicological methods to evaluate petroleum contaminants have assessed tissue burdens, with fewer assays providing indicators of health or physiology, particularly when contaminant levels are low and chronic. Marine mussels are a ubiquitous and crucial component of the nearshore environment, and new genomic technologies exist to quantify molecular responses of individual mussels to stimuli, including exposure to polycyclic aromatic hydrocarbons (PAHs). We used gene-based assays of exposure and physiological function to assess chronic oil contamination using the Pacific blue mussel, Mytilus trossulus . We developed a diagnostic gene transcription panel to investigate exposure to PAHs and other contaminants and its effects on mussel physiology and health. During 2012–2015, we analyzed mussels from five field sites in western Prince William Sound, Alaska, with varying oil histories from the 1989 Exxon Valdez oil spill, and from three boat harbors in the area. Gene transcription patterns of mussels from harbors were consistent with elevated exposure to PAHs or other contaminants, whereas transcription patterns of mussels sampled from shorelines in areas affected by the oil spill indicated no PAH exposure.

Alaska↗

Expanding the role of reactive transport models in critical zone processes

Models test our understanding of processes and can reach beyond the spatial and temporal scales of measurements. Multi-component Reactive Transport Models (RTMs), initially developed more than three decades ago, have been used extensively to explore the interactions of geothermal, hydrologic, geochemical, and geobiological processes in subsurface systems. Driven by extensive data sets now available from intensive measurement efforts, there is a pressing need to couple RTMs with other community models to explore non-linear interactions among the atmosphere, hydrosphere, biosphere, and geosphere. Here we briefly review the history of RTM development, summarize the current state of RTM approaches, and identify new research directions, opportunities, and infrastructure needs to broaden the use of RTMs. In particular, we envision the expanded use of RTMs in advancing process understanding in the Critical Zone, the veneer of the Earth that extends from the top of vegetation to the bottom of groundwater. We argue that, although parsimonious models are essential at larger scales, process-based models offer tools to explore the highly nonlinear coupling that characterizes natural systems. We present seven testable hypotheses that emphasize the unique capabilities of process-based RTMs for (1) elucidating chemical weathering and its physical and biogeochemical drivers; (2) understanding the interactions among roots, micro-organisms, carbon, water, and minerals in the rhizosphere; (3) assessing the effects of heterogeneity across spatial and temporal scales; and (4) integrating the vast quantity of novel data, including “omics” data (genomics, transcriptomics, proteomics, metabolomics), elemental concentration and speciation data, and isotope data into our understanding of complex earth surface systems. With strong support from data-driven sciences, we are now in an exciting era where integration of RTM framework into other community models will facilitate process understanding across disciplines and across scales.

Earth-Science Reviews↗