We demonstrate that native grass species from coastal and geothermal habitats require symbiotic fungal endophytes for salt and heat tolerance, respectively. Symbiotically conferred stress tolerance is a habitat-specific phenomenon with geothermal endophytes conferring heat but not salt tolerance, and coastal endophytes conferring salt but not heat tolerance. The same fungal species isolated from plants in habitats devoid of salt or heat stress did not confer these stress tolerances. Moreover, fungal endophytes from agricultural crops conferred disease resistance and not salt or heat tolerance. We define habitat-specific, symbiotically-conferred stress tolerance as habitatadapted symbiosis and hypothesize that it is responsible for the establishment of plants in highstress habitats. The agricultural, coastal and geothermal plant endophytes also colonized tomato (a model eudicot) and conferred disease, salt and heat tolerance, respectively. In addition, the coastal plant endophyte colonized rice (a model monocot) and conferred salt tolerance. These endophytes have a broad host range encompassing both monocots and eudicots. Interestingly, the endophytes also conferred drought tolerance to plants regardless of the habitat of origin. Abiotic stress tolerance correlated either with a decrease in water consumption or reactive oxygen sensitivity/generation but not to increased osmolyte production. The ability of fungal endophytes to confer stress tolerance to plants may provide a novel strategy for mitigating the impacts of global climate change on agricultural and native plant communities.
Environmental DNA (eDNA) has emerged as a potentially powerful tool for use in conservation and resource management, including for tracking the recolonization dynamics of fish populations. We used eDNA to assess the effectiveness of dam removal to restore fish passage on the Elwha River in Washington State (USA). Using a suite of 11 species‐specific eDNA polymerase chain reaction (PCR) assays, we showed that most targeted anadromous species (five Pacific Salmon species and Pacific Lamprey) were able to pass upstream of both former dam sites. Multiscale occupancy modeling showed that the timing and spatial extent of recolonization differed among species during the four years of post‐dam removal monitoring. More abundant species like Chinook Salmon and Coho Salmon migrated farther into the upper portions of the watershed than less abundant species like Pink Salmon and Chum Salmon. Sampling also allowed assessment of potamodromous fish species. Bull Trout and Rainbow Trout, ubiquitous species in the watershed, were detected at all sampling locations. Environmental DNA from Brook Trout, a non‐native species isolated between the dams prior to dam removal, was detected downstream of Elwha dam but rarely upstream of the Glines Canyon Dam suggested that the species has not expanded its range appreciably in the watershed following dam removal. We found that eDNA was an effective tool to assess the response of fish populations to large‐scale dam removal on the Elwha River.
Accurate species identification is vital to conservation and management of species at risk. Species identification is challenging when taxa express similar phenotypic characters and form hybrids, for example the endangered shortnose sucker (Chasmistes brevirostris) and Lost River sucker (Deltistes luxatus). Here, we developed 20 Taqman assays that differentiate these species (19 nuclear DNA and one mitochondrial DNA). Assays were evaluated in 160 young-of-the-year identified to species using meristic counts. Alleles were not fixed between species, but species were highly differentiated (F ST = 0.753, P \ 0.001). The assays developed herein will be a valuable tool for resource managers.
In this study, we examine salinity stress tolerances of two populations of the invasive species New Zealand mud snail Potamopyrgus antipodarum, one population from a high salinity environment in the Columbia River estuary and the other from a fresh water lake. In 1996, New Zealand mud snails were discovered in the tidal reaches of the Columbia River estuary that is routinely exposed to salinity at near full seawater concentrations. In contrast, in their native habitat and throughout its spread in the western US, New Zealand mud snails are found only in fresh water ecosystems. Our aim was to determine whether the Columbia River snails have become salt water adapted. Using a modification of the standard amphipod sediment toxicity test, salinity tolerance was tested using a range of concentrations up to undiluted seawater, and the snails were sampled for mortality at daily time points. Our results show that the Columbia River snails were more tolerant of acute salinity stress with the LC 50 values averaging 38 and 22 Practical Salinity Units for the Columbia River and freshwater snails, respectively. DNA sequence analysis and morphological comparisons of individuals representing each population indicate that they were all P. antipodarum. These results suggest that this species is salt water adaptable and in addition, this investigation helps elucidate the potential of this aquatic invasive organism to adapt to adverse environmental conditions.
Surveys of environmental DNA (eDNA) have become an important and multifaceted tool for monitoring and identifying distributions and occupancy of aquatic species. This tool is attractive because it is powerful, easy to apply, and provides an alternative to traditional field survey methods. However, validating eDNA survey methods against traditional field survey methods is warranted prior to their application. We used eDNA and electrofishing to survey 10 sites in 3 tributaries of the Chehalis River, Washington, to infer distribution and occupancy of Entosphenus tridentatus and Lampetra spp. Both methods produced similar detection rates for E. tridentatus, and Lampetra spp. were detected at slightly greater frequency with eDNA in the Black River and Skookumchuck River. Within each of the three tributaries, eDNA concentration was negatively related to sample distance from the Chehalis River mainstem for E. tridentatus but not for Lampetra spp., which indicates E. tridentatus and Lampetra spp. may be distributed differently within tributaries. Application of lamprey eDNA data to a multiscale occupancy model indicated high probability of detecting eDNA in water samples and quantitative PCR (qPCR) assays. Broad distribution and high detection of E. tridentatus and Lampetra spp. suggest robust populations inhabit the Chehalis River basin. Our findings suggest eDNA surveys may be comparable to electrofishing for informing lamprey occupancy and distributions. Such sampling is efficient and cost‐effective and we anticipate that eDNA surveys will become a valuable tool in addressing key research and monitoring needs for conservation and restoration of lampreys in general.
A 3-primer PCR system was developed to discriminate invasive zebra (Dreissena polymorpha) and quagga (Dreissena bugensis) mussel. The system is based on: 1) universal primers that amplifies a region of the nuclear 28s rDNA gene from both species and 2) a species-specific primer complementary to either zebra or quagga mussel. The species-specific primers bind to sequences between the binding sites for the universal primers resulting in the amplification of two products from the target species and one product from the nontarget species. Therefore, nontarget products are positive amplification controls. The 3-primer system accurately discriminated zebra and quagga mussels from seven geographically distinct populations.
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