High levels of gene flow and genetic diversity in Irish populations of Salix caprea L. inferred from chloroplast and nuclear SSR markers

peer-reviewedBackground Salix caprea is a cold-tolerant pioneer species that is ecologically important in Europe and western and central Asia. However, little data is available on its population genetic structure and molecular ecology. We describe the levels of geographic population genetic structure in natural Irish populations of S. caprea and determine the extent of gene flow and sexual reproduction using both chloroplast and nuclear simple sequence repeats (SSRs). Results A total of 183 individuals from 21 semi-natural woodlands were collected and genotyped. Gene diversity across populations was high for the chloroplast SSRs (H T  = 0.21-0.58) and 79 different haplotypes were discovered, among them 48% were unique to a single individual. Genetic differentiation of populations was found to be between moderate and high (mean G ST  = 0.38). For the nuclear SSRs, G ST was low at 0.07 and observed heterozygosity across populations was high (H O  = 0.32-0.51); only 9.8% of the genotypes discovered were present in two or more individuals. For both types of markers, AMOVA showed that most of the variation was within populations. Minor geographic pattern was confirmed by a Bayesian clustering analysis. Gene flow via pollen was found to be approximately 7 times more important than via seeds. Conclusions The data are consistent with outbreeding and indicate that there are no significant barriers for gene flow within Ireland over large geographic distances. Both pollen-mediated and seed-mediated gene flow were found to be high, with some of the populations being more than 200 km apart from each other. These findings could simply be due to human intervention through seed trade or accidental transportation of both seeds and pollen. These results are of value to breeders wishing to exploit natural genetic variation and foresters having to choose planting material.Teagasc Walsh Fellowship Programm

Plastid genome sequencing reveals biogeographical structure and extensive population genetic variation in wild populations of Phalaris arundinacea L. in north‐western Europe

peer-reviewedNew and comprehensive collections of the perennial rhizomatous reed canary grass (Phalaris arundinacea) were made in NW Europe along north‐to‐south and east‐to‐west clines from Denmark, Germany, Ireland, Poland, Sweden and the United Kingdom. Rhizome, seed and leaf samples were taken for analysis and genetic resource conservation. A subsample covering the geographical range was characterized using plastid genome sequencing and SNP discovery generated using a long‐read PCR and MiSeq sequencing approach. Samples were also subject to flow cytometry and all found to be tetraploid. New sequences were assembled against a Lolium perenne (perennial ryegrass) reference genome, and an average of approximately 60% of each genome was aligned (81 064 bp). Genetic variation was high among the 48 sequenced genotypes with a total of 1793 SNPs, equating to 23 SNPs per kbp. SNPs were subject to principal coordinate and Structure analyses to detect population genetic groupings and to examine phylogeographical pattern. Results indicate substantial genetic variation and population genetic structuring of this allogamous species at a broad geographical scale in NW Europe with plastid genetic diversity organized more across an east‐to‐west than a north‐to‐south cline

Genotyping by Sequencing and Plastome Analysis Finds High Genetic Variability and Geographical Structure in Dactylis glomerata L. in Northwest Europe Despite Lack of Ploidy Variation

peer-reviewedLarge collections of the forage and bioenergy grass Dactylis glomerata were made in northwest (NW) Europe along east to west and north to south clines for genetic resource conservation and to inform breeding programmes of genetic diversity, genepools, and ploidy. Leaves were sampled for genetic analysis and seed and rhizome for ex-situ conservation. Genotyping by sequencing (GBS) was used to assay nuclear DNA diversity and plastome single nucleotide polymorphism (SNP) discovery was undertaken using a long-read PCR and MiSeq approach. Nuclear and plastid SNPs were analysed by principal component analysis (PCA) to compare genotypes. Flow cytometry revealed that all samples were tetraploid, but some genome size variation was recorded. GBS detected an average of approximately 10,000 to 15,000 SNPs per country sampled. The highest average number of private SNPs was recorded in Poland (median ca. 2000). Plastid DNA variation was also high (1466 SNPs, 17 SNPs/kbp). GBS data, and to a lesser extent plastome data, also show that genetic variation is structured geographically in NW Europe with loose clustering matching the country of plant origin. The results reveal extensive genetic diversity and genetic structuring in this versatile allogamous species despite lack of ploidy variation and high levels of human mediated geneflow via planting

Variation in sequences containing microsatellite motifs in the perennial biomass and forage grass, Phalaris arundinacea (Poaceae)

This study has been financed under the collaborative European Community Framework FP7 project GrassMargins (KBBE-2011-5-289461).peer-reviewedForty three microsatellite markers were developed for further genetic characterisation of a forage and biomass grass crop, for which genomic resources are currently scarce. The microsatellite markers were developed from a normalized EST-SSR library. All of the 43 markers gave a clear banding pattern on 3 % Metaphor agarose gels. Eight selected SSR markers were tested in detail for polymorphism across eleven DNA samples of large geographic distribution across Europe. The new set of 43 SSR markers will help future research to characterise the genetic structure and diversity of Phalaris arundinacea, with a potential to further understand its invasive character in North American wetlands, as well as aid in breeding work for desired biomass and forage traits. P. arundinacea is particularly valued in the northern latitude as a crop with high biomass potential, even more so on marginal lands.European Community Framework FP

The Juncaceae-Cyperaceae Interface: A Combined Plastid Sequence Analysis

Phylogenetic relationships of Juncaceae and related families of Cyperales were investigated using DNA sequence data from the plastid rps16 intron, trnL intron, and trnL–F intergenic spacer. Results using parsimony analysis of the aligned matrices find Juncaceae and Cyperaceae to be monophyletic families, which form a clade sister to a monophyletic Thurniaceae that includes Prionium (Prioniaceae), all three clades with 100% bootstrap support. Within Juncaceae, the genus Luzula is monophyletic with 100% bootstrap support and sister to the rest of Juncaceae. Further groupings within the family indicate that the genus Juncus may be monophyletic only with the inclusion of the single-flowered genera of Juncaceae (Distichia, Oxychloe, Marsippospermum, and Rostkovia; sequences of Patosia were not included). Major groupings within Juncus are supported by the morphological characters of septate or non-septate leaves and the presence or absence of bracts subtending the flowers, which have been used to define subgenera and sections within Juncus

Large Trees, Supertrees, and Diversification of the Grass Family

Phylogenetic studies of grasses (Poaceae) are advanced in comparison with most other angiosperm families. However, few studies have attempted to build large phylogenetic trees of the family and use these for evaluating patterns of diversification or other macroevolutionary hypotheses. Two contrasting approaches can be used to generate large trees: supermatrix analyses and supertrees. In this paper, we evaluated the suitability of each of these methods for the study of patterns and processes of evolution in the grasses. We collected data from DDBJ/EMBL/GenBank to determine sequence availability and asked how far we are from a complete generic-level phylogenetic tree of the grasses. We generated almost complete tribal-level supertrees (39 tribes) with over 400 genera using MRP methods, described their major clades, assessed their accuracy, and used them for the study of diversification. We generated a proportional supertree, by modifying the original supertree, to remove sampling bias associated with the original supertree that may affect diversification statistics. We used methods that incorporate information on the topological distribution of taxon diversity from all internal nodes of the phylogenetic tree to show that the grasses have experienced significant variations in diversification rates (M statistic P-value

Environmental risk assessment of genetically modified crops: The use of molecular markers to trace insect and wind dispersal of Brassica napus pollen.

End of Project ReportThis project aimed to develop a better understanding of the potential risks of gene flow and to generate isolation distances required for genetically modified (GM) OSR. The study examined biotic (insect) and abiotic (wind) pollen dispersal over two seasons. A considerable portion of work was devoted to the development of molecular markers, to differentiate Brassica napus varieties from each other to distinguish them from their wild relatives. The project broadly aimed to study gene flow, via pollen movement, from OSR but specifically aimed to: ! Characterise B. napus cv. Marinka using molecular markers. ! Elucidate the distance travelled by OSR pollen by biotic dispersal. ! Elucidate the distance travelled by OSR pollen by abiotic dispersal. ! Elucidate pollination/seed set at various distances from a source crop using male sterile bait plants. ! Develop risk assessment/containment strategies

First Report of the Ash Sawfly, Tomostethus nigritus, Established on Fraxinus excelsior in the Republic of Ireland

Publication history: Accepted - 15 December 2021; Published - 21 December 2021.Simple Summary: Ash sawfly, Tomostethus nigritus, larvae feed on the leaves of Fraxinus excelsior. In the last 20 years, outbreaks of these insects have become more common, and they cause severe defoliation of the tree crown. This pest is native to Europe, and it was recorded for the first time on the island of Ireland in Belfast, Northern Ireland (UK) in 2016. This article is the first report of T. nigritus in the Republic of Ireland (IE). Damaged leaves and larvae were observed on ash trees in Co. Kildare in summer 2021. The larvae were collected and then identified using molecular analyses. Similar damaged leaves were observed on trees in Co. Dublin, which showed mild defoliation of the crown. However, severe defoliation of ash trees has also been recorded in Co. Meath in 2021. No control treatments are available against the insects, and little is known about the cause of the outbreaks. Severe outbreaks by T. nigritus could further affect the delicate situation that the Irish ash tree population is currently facing caused by another severe antagonist of Fraxinus, the ash dieback pathogen Hymenoscyphus fraxineus. Abstract: This is the first report of the ash sawfly, Tomostethus nigritus, in the Republic of Ireland. We observed defoliated leaves of Fraxinus excelsior L. and T. nigritus larvae at a forestry plantation in Co. Kildare. Morphological observation of the larvae and DNA analysis using mitochondrial COI barcoding confirmed the identification of this pest of ash.This research was funded by DAFM and DAERA grant number 2019R578, Nexcelsior: Next Steps in managing the impact of ash dieback project

Genomic prediction of crown rust resistance in Lolium perenne

peer-reviewedBackground Genomic selection (GS) can accelerate genetic gains in breeding programmes by reducing the time it takes to complete a cycle of selection. Puccinia coronata f. sp lolli (crown rust) is one of the most widespread diseases of perennial ryegrass and can lead to reductions in yield, persistency and nutritional value. Here, we used a large perennial ryegrass population to assess the accuracy of using genome wide markers to predict crown rust resistance and to investigate the factors affecting predictive ability. Results Using these data, predictive ability for crown rust resistance in the complete population reached a maximum of 0.52. Much of the predictive ability resulted from the ability of markers to capture genetic relationships among families within the training set, and reducing the marker density had little impact on predictive ability. Using permutation based variable importance measure and genome wide association studies (GWAS) to identify and rank markers enabled the identification of a small subset of SNPs that could achieve predictive abilities close to those achieved using the complete marker set. Conclusion Using a GWAS to identify and rank markers enabled a small panel of markers to be identified that could achieve higher predictive ability than the same number of randomly selected markers, and predictive abilities close to those achieved with the entire marker set. This was particularly evident in a sub-population characterised by having on-average higher genome-wide linkage disequilibirum (LD). Higher predictive abilities with selected markers over random markers suggests they are in LD with QTL. Accuracy due to genetic relationships will decay rapidly over generations whereas accuracy due to LD will persist, which is advantageous for practical breeding applications.This work received funding from the Irish Department of Agriculture Food and the Marine DAFM (RSF 11/S/109) and Teagasc core funding. SKA is supported by a Teagasc PhD Walsh Fellowship. SLB has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No. 658031

Phylogenetics of Taxus Using the Internal Transcribed Spacers of Nuclear Ribosomal DNA and Plastid trnL-F Regions

Taxus is a genus of trees and shrubs with high value in horticulture and medicine as a source of the anticancer drug paclitaxel. The taxonomy of the group is complex due to the lack of diagnostic morphological characters and the high degree of similarity among species. Taxus has a wide global geographic distribution and some taxonomists recognize only a single species with geographically defined subgroups, whereas others have described several species. To address these differences in taxonomic circumscription, phylogenetic analyses were conducted on DNA sequences using Maximum Likelihood, Bayesian Inference and TCS haplotype networks on single and combined gene regions obtained for the nuclear ribosomal ITS region and the plastid trnL intron and trnL-F intergenic spacer. Evidence is presented for the sister group status of Pseudotaxus to Taxus and the inclusion of Amentotaxus, Austrotaxus, Cephalotaxus and Torreya within Taxaceae. Results are consistent with the taxonomic recognition of nine species: T. baccata, T. brevifolia, T. canadensis, T. cuspidata, T. floridana, T. fuana, T. globosa, T. sumatrana and T. wallichiana, but evidence is found for less species distinction and considerable reticulation within the T. baccata, T. canadensis and T. cuspidata group. We compare the results to known taxonomy, biogeography, present new leaf anatomical data and discuss the origins of the hybrids T. ×media and T. ×hunnewelliana