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1. On the potential of Angiosperms353 for population genomic studies

   https://doi.org/10.1002/aps3.11419  

   Slimp, Madeline ; Williams, Lindsay D. ; Hale, Haley ; Johnson, Matthew G. ( June 2021 , Applications in Plant Sciences)

   null (Ed.)

   PREMISE The successful application of universal targeted sequencing markers, such as those developed for the Angiosperms353 probe set, within populations could reduce or eliminate the need for specific marker development, while retaining the benefits of full-gene sequences in population-level analyses. However, whether the Angiosperms353 markers provide sufficient variation within species to calculate demographic parameters is untested. METHODS Using herbarium specimens from a 50-year-old floristic survey in Texas, we sequenced 95 samples from 24 species using the Angiosperms353 probe set. Our data workflow calls variants within species and prepares data for population genetic analysis using standard metrics. In our case study, gene recovery was affected by genomic library concentration only at low concentrations and displayed limited phylogenetic bias. RESULTS We identified over 1000 segregating variants with zero missing data for 92% of species and demonstrate that Angiosperms353 markers contain sufficient variation to estimate pairwise nucleotide diversity (π)—typically between 0.002 and 0.010, with most variation found in flanking non-coding regions. In a subset of variants that were filtered to reduce linkage, we uncovered high heterozygosity in many species, suggesting that denser sampling within species should permit estimation of gene flow and population dynamics. DISCUSSION Angiosperms353 should benefit conservation genetic studies by providing universal repeatable markers, low missing data, and haplotype information, while permitting inclusion of decades-old herbarium specimens. 

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2. Incomplete reproductive isolation and low genetic differentiation despite floral divergence across varying geographic scales in Castilleja

   https://doi.org/10.1002/ajb2.1700  

   Wenzell, Katherine E. ; McDonnell, Angela J. ; Wickett, Norman J. ; Fant, Jeremie B. ; Skogen, Krissa A. ( July 2021 , American Journal of Botany)

   Divergence depends on the strength of selection and frequency of gene flow between taxa, while reproductive isolation relies on mating barriers and geographic distance. Less is known about how these processes interact at early stages of speciation. Here, we compared population‐level differentiation in floral phenotype and genetic sequence variation among recently divergedCastillejato explore patterns of diversification under different scenarios of reproductive isolation.

   Using target enrichment enabled by the Angiosperms353 probe set, we assessed genetic distance among 50 populations of fourCastillejaspecies. We investigated whether patterns of genetic divergence are explained by floral trait variation or geographic distance in two focal groups: the widespreadC. sessilifloraand the more restrictedC. purpureaspecies complex.

   We document thatC. sessilifloraand theC. purpureacomplex are characterized by high diversity in floral color across varying geographic scales. Despite phenotypic divergence, groups were not well supported in phylogenetic analyses, and little genetic differentiation was found across targeted Angiosperms353 loci. Nonetheless, a principal coordinate analysis of single nucleotide polymorphisms revealed differentiation withinC. sessilifloraacross floral morphs and geography and less differentiation among species of theC. purpureacomplex.

   Patterns of genetic distance inC. sessiliflorasuggest species cohesion maintained over long distances despite variation in floral traits. In theC. purpureacomplex, divergence in floral color across narrow geographic clines may be driven by recent selection on floral color. These contrasting patterns of floral and genetic differentiation reveal that divergence can arise via multiple eco‐evolutionary paths.

    

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3. The Sticta filix - Sticta lacera conundrum (lichenized Ascomycota: Peltigeraceae subfamily Lobarioideae): unresolved lineage sorting or developmental switch?

   https://doi.org/10.1093/botlinnean/boab083  

   Lücking, Robert ; Moncada, Bibiana ; Widhelm, Todd_J ; Lumbsch, H_Thorsten ; Blanchon, Dan_J ; de_Lange, Peter_J ( December 2021 , Botanical Journal of the Linnean Society)

   We assessed the status of two New Zealand endemic morphodemes in the genus Sticta, currently treated as two separate taxa, Sticta filix and Sticta lacera. Both are green-algal lichens with a distinct stipe that grow in forested habitats and are suitable indicators of the indigenous vegetation health in forest ecosystems in New Zealand. They exhibit different morphologies and substrate ecologies: S. filix forms rather robust thalli, often on exposed trunks of phorophytes, with erect stems distinctly emerging from the substrate, whereas S. lacera is a more delicate lichen growing near the base of trees, usually among bryophyte mats or sheltered in the exposed portions of the phorophyte root-plate, with a prostrate, branched, stolon-like stem barely emerging from the substrate. Throughout their range, both taxa grow sympatrically and often in close proximity (syntopically). Despite the differences, ITS barcoding does not support the two morphodemes as separate species. In this study we assessed two possible explanations: (1) S. filix and S. lacera are discrete phenotypes of a single species, caused by developmental switching triggered by a discrete environmental variable, the propagules developing either on bare substrate or between bryophytes; and (2) the two morphodemes represent separate lineages, but ITS does not provide sufficient resolution to reflect this. We performed a quantitative analysis of morphological and ecological parameters, based on vouchered herbarium collections and field observations on iNaturalist NZ (https://inaturalist.nz), to assess the level of discreteness of the growth forms and to test for a correlation with the presence of a bryophyte mat. We further took advantage of an existing molecular data set from a target capture approach, comprised of 205 protein markers. This data set was used to establish a framework of percentage identities between pairs of the same and of different species among lobarioid Peltigeraceae and then to test whether the S. filix/lacera pairing fell closer to a within-species or a between-species pairing. The morphometric analysis of herbarium material resolved S. filix and S. lacera as two discrete morphs with little overlap, supported by numerous observations on iNaturalist NZ. However, whereas herbarium material suggested a significant association of the lacera morph with bryophyte mats, no such pattern was evident from field images on iNaturalist NZ, in which both morphs frequently associated with bryophyte mats. This highlights the limitations of herbarium material to correctly assess substrate ecology, whereas iNaturalist NZ postings had issues with correct identifications, given that especially S. lacera is easily confused with Pseudocyphellaria multifida. Based on the target capture data, the percentage identity of the S. filix/lacera pairing (99.43%) was significantly higher than that of all 12 between-species pairings (93.20–98.01%); it was at the same time lower than that of all within-species pairings (99.63–99.99%) but significantly so only in comparison with five out of the eight within-species pairings. The target capture approach is thus inconclusive, but the combination of all data suggests that S. filix and S. lacera are not discrete morphodemes of a single species but represent two separate lineages which emerged recently and hence cannot be resolved using the ITS barcoding marker or even a deeper phylogenomic approach based on protein-coding markers. We propose transplantation experiments and the application of RADseq to further assess this situation.

    

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4. Lineage‐specific vs. universal: A comparison of the Compositae1061 and Angiosperms353 enrichment panels in the sunflower family

   https://doi.org/10.1002/aps3.11422  

   Siniscalchi, Carolina M. ; Hidalgo, Oriane ; Palazzesi, Luis ; Pellicer, Jaume ; Pokorny, Lisa ; Maurin, Olivier ; Leitch, Ilia J. ; Forest, Felix ; Baker, William J. ; Mandel, Jennifer R. ( June 2021 , Applications in Plant Sciences)

   Phylogenetic studies in the Compositae are challenging due to the sheer size of the family and the challenges they pose for molecular tools, ranging from the genomic impact of polyploid events to their very conserved plastid genomes. The search for better molecular tools for phylogenetic studies led to the development of the family‐specific Compositae1061 probe set, as well as the universal Angiosperms353 probe set designed for all flowering plants. In this study, we evaluate the extent to which data generated using the family‐specific kit and those obtained with the universal kit can be merged for downstream analyses.

   We used comparative methods to verify the presence of shared loci between probe sets. Using two sets of eight samples sequenced with Compositae1061 and Angiosperms353, we ran phylogenetic analyses with and without loci flagged as paralogs, a gene tree discordance analysis, and a complementary phylogenetic analysis mixing samples from both sample sets.

   Our results show that the Compositae1061 kit provides an average of 721 loci, with 9–46% of them presenting paralogs, while the Angiosperms353 set yields an average of 287 loci, which are less affected by paralogy. Analyses mixing samples from both sets showed that the presence of 30 shared loci in the probe sets allows the combination of data generated in different ways.

   Combining data generated using different probe sets opens up the possibility of collaborative efforts and shared data within the synantherological community.

    

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5. A basic dd RAD seq two‐enzyme protocol performs well with herbarium and silica‐dried tissues across four genera

   https://doi.org/10.1002/aps3.11344  

   Jordon‐Thaden, Ingrid E. ; Beck, James B. ; Rushworth, Catherine A. ; Windham, Michael D. ; Diaz, Nicolas ; Cantley, Jason T. ; Martine, Christopher T. ; Rothfels, Carl J. ( April 2020 , Applications in Plant Sciences)

   The ability to sequence genome‐scale data from herbarium specimens would allow for the economical development of data sets with broad taxonomic and geographic sampling that would otherwise not be possible. Here, we evaluate the utility of a basic double‐digest restriction site–associatedDNAsequencing (ddRADseq) protocol usingDNAs from four genera extracted from both silica‐dried and herbarium tissue.

   DNAs fromDraba,Boechera,Solidago, andIlexwere processed with a ddRADseq protocol. The effects ofDNAdegradation, taxon, and specimen age were assessed.

   Although taxon, preservation method, and specimen age affected data recovery, large phylogenetically informative data sets were obtained from the majority of samples.

   These results suggest that herbarium samples can be incorporated into ddRADseq project designs, and that specimen age can be used as a rapid on‐site guide for sample choice. The detailed protocol we provide will allow users to pursue herbarium‐based ddRADseq projects that minimize the expenses associated with fieldwork and sample evaluation.

    

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