Kevin Weitemier’s research while affiliated with Oregon State University and other places

Aim Biodiversity changes can occur gradually or as distinct jumps as species distributions end and others begin. However, comprehensive assessments of freshwater biodiversity and how assemblages change along a continuum have been hindered by the connectivity and networked nature of streams and their large numbers of rare and cryptic species found there. We hypothesize that the boundary where fish distributions end marks an ecological shift in freshwater assemblages and could unify stream concepts. Location Trask River watershed, Oregon, USA, temperate North America. Taxon Fishes, amphibians, crayfishes, bivalves, mammals, pathogens. Methods We targeted freshwater species across taxa using multigene eDNA metabarcoding of 48 different DNA barcodes to identify assemblages to understand species patterns distributed along a longitudinal gradient in watersheds around the upper fish distribution boundary. Detections from multiple genetic sources improve the detection probability for rare species. Results We detected a shift in the assemblage from lower reaches towards headwaters where fish distributions end, leading to fewer species across taxa at reduced eDNA signal. We uncovered a hidden biodiversity in watersheds by detecting a sparsely distributed amphibian pathogen Batrachochytrium dendrobatidis , the elusive mammal Mountain Beaver, fish species that went undetected using traditional approaches, and newly discovered cryptic lineages of sculpins. Salmonids were detected further upstream with eDNA than expected. Main conclusions Our findings unify stream concepts owing to the marked species shift at the upper extent of fish elevating the importance of this boundary. The detection of salmonids further upstream suggests a distribution extension. The sculpin detections reveal a cryptic species complex with ramifications for fish conservation, potential pockets of endemism and possible identification of new species. We show extensive diversity in watersheds, some of which was previously hidden, transforming our understanding of species presence and distributions affecting our ability to track, protect and manage them across watersheds.

Background Hawthorn species ( Crataegus L.; Rosaceae tribe Maleae) form a well-defined clade comprising five subgeneric groups readily distinguished using either molecular or morphological data. While multiple subsidiary groups (taxonomic sections, series) are recognized within some subgenera, the number of and relationships among species in these groups are subject to disagreement. Gametophytic apomixis and polyploidy are prevalent in the genus, and disagreement concerns whether and how apomictic genotypes should be recognized taxonomically. Recent studies suggest that many polyploids arise from hybridization between members of different infrageneric groups. Methods We used target capture and high throughput sequencing to obtain nucleotide sequences for 257 nuclear loci and nearly complete chloroplast genomes from a sample of hawthorns representing all five currently recognized subgenera. Our sample is structured to include two examples of intersubgeneric hybrids and their putative diploid and tetraploid parents. We queried the alignment of nuclear loci directly for evidence of hybridization, and compared individual gene trees with each other, and with both the maximum likelihood plastome tree and the nuclear concatenated and multilocus coalescent-based trees. Tree comparisons provided a promising, if challenging (because of the number of comparisons involved) method for visualizing variation in tree topology. We found it useful to deploy comparisons based not only on tree-tree distances but also on a metric of tree-tree concordance that uses extrinsic information about the relatedness of the terminals in comparing tree topologies. Results We obtained well-supported phylogenies from plastome sequences and from a minimum of 244 low copy-number nuclear loci. These are consistent with a previous morphology-based subgeneric classification of the genus. Despite the high heterogeneity of individual gene trees, we corroborate earlier evidence for the importance of hybridization in the evolution of Crataegus . Hybridization between subgenus Americanae and subgenus Sanguineae was documented for the origin of Sanguineae tetraploids, but not for a tetraploid Americanae species. This is also the first application of target capture probes designed with apple genome sequence. We successfully assembled 95% of 257 loci in Crataegus , indicating their potential utility across the genera of the apple tribe.

Genetic diversity underpins species conservation and management goals, and ultimately determines a species' ability to adapt. Using freshwater environmental DNA (eDNA) samples, we examined mitochondrial genetic diversity using multigene metabarcode sequence data from four Oncorhynchus species across 16 sites in Oregon and northern California. Our multigene metabarcode panel included targets commonly used in population genetic NADH dehydrogenase 2 (ND2), phylogenetic cytochrome c oxidase subunit 1 (COI) and eDNA (12S ribosomal DNA) screening. The ND2 locus showed the greatest within-species haplotype diversity for all species, followed by COI and then 12S rDNA for all species except Oncorhynchus kisutch. Sequences recovered for O. clarkii clarkii were either identical to, or one mutation different from, previously characterized haplotypes (95.3% and 4.5% of reads, respectively). The greatest diversity in O. c. clarkii was among coastal watersheds, and subsets of this diversity were shared with fish in inland watersheds. However, coastal streams and the Umpqua River watershed appear to harbour unique haplotypes. Sequences from O. mykiss revealed a disjunction between the Willamette watershed and southern watersheds suggesting divergent histories. We also identified similarities between populations in the northern Deschutes and southern Klamath watersheds, consistent with previously hypothesized connections between the two via inland basins. Oncorhynchus kisutch was only identified in coastal streams and the Klamath River watershed, with most diversity concentrated in the coastal Coquille watershed. Oncorhynchus tshawytscha was only observed at one site, but contained multiple haplotypes at each locus. The characterization of genetic diversity at multiple loci expands the knowledge gained from eDNA sampling and provides crucial information for conservation actions and genetic management.

Taxonomic monographs have the potential to make a unique contribution to the understanding of global biodiversity. However, such studies, now rare, are often considered too daunting to undertake within a realistic time frame, especially as the world’s collections have doubled in size in recent times. Here, we report a global-scale monographic study of morning glories (Ipomoea) that integrated DNA barcodes and high-throughput sequencing with the morphological study of herbarium specimens. Our approach overhauled the taxonomy of this megadiverse group, described 63 new species and uncovered significant increases in net diversification rates comparable to the most iconic evolutionary radiations in the plant kingdom. Finally, we show that more than 60 species of Ipomoea, including sweet potato, independently evolved storage roots in pre-human times, indicating that the storage root is not solely a product of human domestication but a trait that predisposed the species for cultivation. This study demonstrates how the world’s natural history collections can contribute to global challenges in the Anthropocene. Taxonomic monographs have been considered too vast and daunting as a source for studying biodiversity, but this novel study of morning glories combines herbarium specimens with DNA barcodes and high-throughput sequencing to describe new species and discover hidden traits.

Milkweeds ( Asclepias ) are used in wide-ranging studies including floral development, pollination biology, plant-insect interactions and co-evolution, secondary metabolite chemistry, and rapid diversification. We present a transcriptome and draft nuclear genome assembly of the common milkweed, Asclepias syriaca . This reconstruction of the nuclear genome is augmented by linkage group information, adding to existing chloroplast and mitochondrial genomic resources for this member of the Apocynaceae subfamily Asclepiadoideae. The genome was sequenced to 80.4× depth and the draft assembly contains 54,266 scaffolds ≥1 kbp, with N50 = 3,415 bp, representing 37% (156.6 Mbp) of the estimated 420 Mbp genome. A total of 14,474 protein-coding genes were identified based on transcript evidence, closely related proteins, and ab initio models, and 95% of genes were annotated. A large proportion of gene space is represented in the assembly, with 96.7% of Asclepias transcripts, 88.4% of transcripts from the related genus Calotropis , and 90.6% of proteins from Coffea mapping to the assembly. Scaffolds covering 75 Mbp of the Asclepias assembly formed 11 linkage groups. Comparisons of these groups with pseudochromosomes in Coffea found that six chromosomes show consistent stability in gene content, while one may have a long history of fragmentation and rearrangement. The progesterone 5β-reductase gene family, a key component of cardenolide production, is likely reduced in Asclepias relative to other Apocynaceae. The genome and transcriptome of common milkweed provide a rich resource for future studies of the ecology and evolution of a charismatic plant family.

These are the Abstracts of the 2019 annual meeting of SNVB, WA TWS, and NW PARC. I have two abstracts in this compilation: Olson et al., Density Management and Riparian Buffer Study update, pp 154-155; Weil and Olson, NW PARC update, pp 162-163.

Environmental DNA (eDNA) assays for single‐ and multi‐species detection show promise for providing standardized assessment methods for diverse taxa, but techniques for evaluating multiple taxonomically divergent assemblages are in their infancy. We evaluated whether microfluidic multiplex metabarcoding on the Fluidigm Access Array™ platform and high‐throughput sequencing could identify diverse stream and riparian assemblages from 48 taxon‐general and taxon‐specific metabarcode primers. eDNA screening was paired with electrofishing along a stream continuum to evaluate congruence between methods. A fish hatchery located midway along the stream continuum provided a dispersal barrier, and a point source for non‐native White Sturgeon (Acipencer transmontanus). Microfluidic metabarcoding had 87% accuracy with respect to electrofishing and detected all 13 species electrofishing observed. Taxon‐specific barcoding primers were more successful than taxon‐general universal metabarcoding primers at classifying sequences to species. Both types of markers detected a transition from downstream sites dominated by multiple fish species, to upstream sites dominated by a single species; however, we failed to detect a complementary transition in amphibian occupancy. White Sturgeon was only detected at the hatchery outflow, indicating eDNA transport was not detectable ~2.4 km from its source. Overall, we identified 878 predicted taxa. Most sequences (50.1%) derived from fish (Actinopteri, Petromyzontidae), oomycetes (21.3%), arthropoda (classes Insecta, Decapoda; 16.5%), and apicomplexan parasites (3.8%). Taxa accounting for ~1% or less of sequences included freshwater red algae, diatoms, amphibians, and beaver. Our work shows that microfluidic metabarcoding can survey multiple phyla per assay, providing fine discrimination required to resolve closely related species, and enable data‐driven prioritization for multiple forest health objectives.

Milkweeds ( Asclepias ) are used in wide-ranging studies including floral development, pollination biology, plant-insect interactions and co-evolution, secondary metabolite chemistry, and rapid diversification. We present a transcriptome and draft nuclear genome assembly of the common milkweed, Asclepias syriaca . This reconstruction of the nuclear genome is augmented by linkage group information, adding to existing chloroplast and mitochondrial genomic resources for this member of the Apocynaceae subfamily Asclepiadoideae. The genome was sequenced to 80.4× depth and the draft assembly contains 54,266 scaffolds ≥1 kbp, with N50 = 3415 bp, representing 37% (156.6 Mbp) of the estimated 420 Mbp genome. A total of 14,474 protein-coding genes were identified based on transcript evidence, closely related proteins, and ab initio models, and 95% of genes were annotated. A large proportion of gene space is represented in the assembly, with 96.7% of Asclepias transcripts, 88.4% of transcripts from the related genus Calotropis , and 90.6% of proteins from Coffea mapping to the assembly. Scaffolds covering 75 Mbp of the Asclepias assembly formed eleven linkage groups. Comparisons of these groups with pseudochromosomes in Coffea found that six chromosomes show consistent stability in gene content, while one may have a long history of fragmentation and rearrangement. The progesterone 5β-reductase gene family, a key component of cardenolide production, is likely reduced in Asclepias relative to other Apocynaceae. The genome and transcriptome of common milkweed provide a rich resource for future studies of the ecology and evolution of a charismatic plant family.

Milkweeds ( Asclepias ) are used in wide-ranging studies including floral development, pollination biology, plant-insect interactions and co-evolution, secondary metabolite chemistry, and rapid diversification. We present a transcriptome and draft nuclear genome assembly of the common milkweed, Asclepias syriaca . This reconstruction of the nuclear genome is augmented by linkage group information, adding to existing chloroplast and mitochondrial genomic resources for this member of the Apocynaceae subfamily Asclepiadoideae. The genome was sequenced to 80.4× depth and the draft assembly contains 54,266 scaffolds ≥1 kbp, with N50 = 3415 bp, representing 37% (156.6 Mbp) of the estimated 420 Mbp genome. A total of 14,474 protein-coding genes were identified based on transcript evidence, closely related proteins, and ab initio models, and 95% of genes were annotated. A large proportion of gene space is represented in the assembly, with 96.7% of Asclepias transcripts, 88.4% of transcripts from the related genus Calotropis , and 90.6% of proteins from Coffea mapping to the assembly. Scaffolds covering 75 Mbp of the Asclepias assembly formed eleven linkage groups. Comparisons of these groups with pseudochromosomes in Coffea found that six chromosomes show consistent stability in gene content, while one may have a long history of fragmentation and rearrangement. The progesterone 5β-reductase gene family, a key component of cardenolide production, is likely reduced in Asclepias relative to other Apocynaceae. The genome and transcriptome of common milkweed provide a rich resource for future studies of the ecology and evolution of a charismatic plant family.