Understanding how host–microbiome interactions influence tree disease is critical for understanding forest resilience. Here, we present foliar microbiome ITS2 metabarcoding and transcriptomic datasets from Pinus sylvestris to investigate susceptibility to Dothistroma needle blight (DNB), a globally important foliar disease caused by Dothistroma septosporum. We hypothesised that host genotype shapes foliar microbial communities and their interactions, thereby influencing disease outcomes. Samples were collected from a progeny–provenance field trial in the south of Scotland representing a broad spectrum of disease susceptibilities. The dataset comprises of ITS2 metabarcoding samples from 200 genotypes across three timepoints and RNAseq samples from 48 genotypes across two timepoints. Sampling captured key stages of pathogen exposure and disease progression. Both standardised and bespoke protocols were used for nucleotide extraction, sequencing, and quality control, including multiple negative and positive controls. These datasets, available in the European Nucleotide Archive (project accession PRJEB88228), enable analysis of temporal dynamics in foliar fungal communities, host–microbiome transcriptional responses, and genotype-dependent variation in disease susceptibility.
Tomato is a staple crop and an excellent model to study host-microbiota interactions in the plant food chain. In this study, we describe a "lab-in-the-field" approach to investigate the microbiota of field-grown tomato plants. High-throughput amplicon sequencing revealed a three-microhabitat partition, phyllosphere, rhizosphere, and root interior, differentiating host-associated communities from the environmental microbiota. An individual bacterium, classified as Acinetobacter sp., emerged as a dominant member of the microbiota at the plant-soil continuum. To gain insights into the functional significance of this enrichment, we subjected rhizosphere specimens to shotgun metagenomics. Similar to the amplicon sequencing survey, a "microhabitat effect," defined by a set of rhizosphere-enriched functions, was identified. Mobilization of mineral nutrients, as well as adaptation to salinity and polymicrobial communities, including antimicrobial resistance genes (ARGs), emerged as a functional requirement sustaining metagenomic diversification. A metagenome-assembled genome representative of Acinetobacter calcoaceticus was retrieved, and metagenomic reads associated with this species identified a functional specialization for plant-growth promotion traits, such as phosphate solubilization, siderophore production, and reactive oxygen species detoxification, which were similarly represented in a tomato genotype-independent fashion. Our results revealed that the enrichment of a beneficial bacterium capable of alleviating plant abiotic stresses appears decoupled from ARGs facilitating microbiota persistence at the root-soil interface.IMPORTANCETomatoes are at center stage in global food security due to their high nutritional value, widespread cultivation, and versatility. Tomatoes provide essential vitamins and minerals, contribute to diverse diets, and support farmer livelihoods, making them a cornerstone of sustainable food systems. Beyond direct dietary benefits, the intricate relationship between tomatoes, their associated microbiota, and antimicrobial resistance gene (ARG) is increasingly recognized. Tomato plants host diverse microbial communities in association with their organs, which influence plant health and productivity. Crop management impacts the composition and function of these communities, contributing to the prevalence of ARGs in the soil and on the plants themselves. These genes can potentially transfer to human pathogens, posing a food safety and public health risk. Understanding these complex interactions is critical for developing sustainable agricultural practices capable of mitigating the impact of climatic modifications and the global threat of antimicrobial resistance.
This article serves as a data repository for recorded crosses made for spring and winter malting barley breeding in Ireland as a collaboration over 100 years between the Department of Agriculture, Food and the Marine (and its precursor organisation) and Guinness. It describes the genotypes and phenotypes of the surviving 37 winter barley breeding lines that are held in the Department of Agriculture, Food and the Marine gene bank and provides a genotypic description of the spring barley breeding lines.This is the first compilation of all surviving records for the crosses made in both spring and winter barley breeding programmes. Breeding for a certain climate and soil conditions will produce excellent varieties, including, in some cases, varieties with a wide European spread such as 'Spratt-Archer'. The Irish malting barley breeding programme was very advanced for its time, incorporating the latest breeding techniques (pedigree breeding, mutation breeding, connectivity to European brewery convention trial network) and advanced quality testing in experimental micro-breweries. The breeding programme has pioneered participatory plant breeding involving the growers early in the selection process.
Iceland was among the last large islands settled by humans, with colonization (Landnám) in the late 9th century CE (Common Era) and is often portrayed as an ecological disaster driven by the Norse settlers. Here, we revisit this narrative through environmental DNA (eDNA) and multiproxy analyses of sediment cores from Lake Tjörnin in central Reykjavík, one of Iceland’s earliest and longest-occupied settlements. Originally a marine embayment, Tjörnin became a freshwater lake around 660 CE. Our record reveals a human presence decades before the long-accepted arrival date of 877 CE, marked by the Landnám volcanic tephra. Early settlement brought livestock, barley cultivation, and other introduced taxa that enhanced nutrient cycling and unexpectedly increased local biodiversity. Contrary to the conventional view of rapid deforestation, eDNA shows that birch and willow expanded during the settlement period, likely supported by deliberate management. Pronounced ecological and land use shifts occurred after 1200 CE, but these were coeval with the Little Ice Age cooling, compounded by volcanic eruptions, storm surges, and plague, rather than anthropogenic degradation. Crop cultivation ceased, arboreal taxa retracted, and grazing pressure maintained open landscapes. Even more profound ecological changes came after c. 1750 CE with urbanization and industrialization, as wastewater discharge, heavy-metal pollution, and fossil fuel use reshaped Tjörnin’s ecosystem. These findings challenge the prevailing model of Norse-induced environmental collapse, revealing instead a dynamic human–environment relationship shaped by both cultural practices and external stressors. By applying eDNA to a long-occupied urban catchment, we demonstrate the power of genomic methods to refine settlement chronologies, reassess ecological baselines and changes, and integrate natural and cultural histories. This approach offers a model for revisiting human–environment interactions in urban centers worldwide. ### Competing Interest Statement The authors have declared no competing interest.
The WHIRLY (WHY) DNA/RNA binding proteins fulfil multiple but poorly characterised functions in leaf development. WHY1 transcript levels were highest in the bases of 7-day old barley leaves. Immunogold labelling revealed that the WHY1 protein was more abundant in the nuclei than the proplastids of the leaf bases. Transcript and metabolite profiling analysis of barley lines (W1-1 and W1-7) lacking WHY1, which show delayed greening compared to the wild type. While the transcript profile of leaf development was largely unchanged in W1-1 and W1-7 leaves, there were differences in levels of several transcripts encoding transcription factors associated with chloroplast development. These include a barley homologue of the Arabidopsis GATA transcription factor that regulates stomatal development, greening and chloroplast development, NAC1, two transcripts with similarity to Arabidopsis GLK1 and two transcripts encoding ARF transcriptions factors with functions in leaf morphogenesis and development. Chloroplast proteins were less abundant in the W1-1 and W1-7 leaves than the wildtype. The levels of TCA cycle metabolites and GABA were significantly lower in WHY1 knockdown leaves than the wild type. We conclude that WHY1 functions in the nuclei of the cells in the leaf bases contributes to the regulation of chloroplast development.
ABSTRACT Winter dormancy is a key process in the phenology of temperate perennials. The changing climate is severely impacting its course leading to economic losses in agriculture. A better understanding of the underlying mechanisms, as well as the genetic basis of the different responses, are necessary for the development of climate-resilient cultivars. This study aims to provide an insight into winter dormancy in red raspberry ( Rubus idaeus L). We report the transcriptomic profiles during dormancy in two raspberry cultivars with contrasting responses. The cultivar ‘Glen Ample’ showed a typical perennial phenology, whereas ‘Glen Dee’ registered consistent dormancy dysregulation, exhibiting active growth and flowering out of season. RNA-seq combined with weighted gene co-expression network analysis (WGCNA) highlighted gene clusters in both genotypes that exhibited time-dependent expression profiles, Functional analysis of ‘Glen Ample’ gene clusters highlighted the significance of the cell and structural development prior to dormancy entry as well the role of genetic and epigenetic processes such as RNAi and DNA methylation in regulating gene expression. On the contrary, dormancy release in ‘Glen Ample’ was associated with upregulation of transcripts associated with the resumption of metabolism, nucleic acid biogenesis and processing signal response pathways. Many of the processes occurring in ‘Glen Ample’ were dysregulated in ‘Glen Dee’ and twenty-eight transcripts exhibiting time-dependent expression in ‘Ample’ that also had an Arabidopsis homologue were not found in all samples from ‘Glen Dee’. These included a gene with homology to Arabidopsis VRN1 ( RiVRN1.1 ) that exhibited a sharp decline in expression following dormancy induction in Glen Ample. Characterisation of the gene region in the ‘Glen Dee’ genome revealed two large insertions upstream of the ATG start codon. We propose that non-expression of a specific VRN1 homologue in ‘Glen Dee’ causes dormancy misregulation as a result of inappropriate expression of a subset of genes that are directly or indirectly regulated by RiVRN1.1 . HIGHLIGHT The raspberry cultivar Glen Dee exhibits aberrant winter dormancy status associated with insertions in the upstream promoter region of a VRN1 ( RiVRN1.1 ) homologue that silence expression, allowing the identification of dormancy-associated genetic modules that are regulated by RiVRN1.1 .
De novo genotyping in potato using methylation-sensitive GBS discovers SNPs largely confined to genic or gene-associated regions and displays enhanced effectiveness in estimating LD decay rates, population structure and detecting GWAS associations over ‘fixed’ SNP genotyping platform. Study also reports the genetic architectures including robust sequence-tagged marker–trait associations for sixteen important potato traits potentially carrying higher transferability across a wider range of germplasm. This study deploys recent advancements in polyploid analytical approaches to perform complex trait analyses in cultivated tetraploid potato. The study employs a ‘fixed’ SNP Infinium array platform and a ‘flexible and open’ genome complexity reduction-based sequencing method (GBS, genotyping-by-sequencing) to perform genome-wide association studies (GWAS) for several key potato traits including the assessment of population structure and linkage disequilibrium (LD) in the studied population. GBS SNPs discovered here were largely confined ( 90
Peat-based growing media products are being phased out in the UK. Alternative constituents may be at risk of harbouring oomycete plant pathogens. Thirty-five peat-free growing media samples were tested for oomycete pathogens by baiting and DNA metabarcoding. No Phytophthora species was detected by baiting, but seven species of Pythium, Phytopythium and Elongisporangium, including four species not previously reported in the UK, were baited. DNA matching eleven Phytophthora species was detected by metabarcoding across a range of samples. A coir sample was also found to contain DNA of Peronosclerospora, a genus of tropical downy mildew pathogens. DNA matching six Phytophthora pathogens, including P. bishii, a pathogen of strawberry not previously reported in the UK, was detected in recycled coir that had been used for strawberry production in The Netherlands. We discuss the implications of the species detected in terms of risks to plant health and the need for further evidence-gathering.
● GERMIN3 has previously been identified as a target of the tuberigen activation complex suggesting a function in potato tuberisation but its role is presently unknown. ● To understand the role of GERMIN3 we analysed morphological, agronomic and molecular phenotypes in transgenic lines. ● GERMIN3 over-expressing lines of Solanum tuberosum ssp. andigena exhibited increased tuber yields under permissive conditions and enhanced tuber numbers. Post-harvest tuber sprouting exhibited reduced apical dominance with increased numbers of sprouts. Apical dominance was reduced in aerial tissues of mature plants where stem growth from axillary buds was activated. Similar results were observed in the commercial cultivar Desiree. Over-expression of GERMIN3 had no impact on the expression of SP6A, a positive regulator of tuberisation or TFL1B, a negative regulator. The GERMIN3 protein localised to the endoplasmic reticulum and transient expression in N. benthamiana leaves resulted in plasmodesmatal gating allowing intercellular transport of GFP-tagged sporamin independent of GERMIN3 oxalate oxidase activity. ● GERMIN3 affects tuberisation and other developmental processes by facilitating meristem activation. This identifies GERMIN3 as a novel protein associated with control of plasmodesmatal transport and supports the importance of plasmodesmatal gating in the regulation of key potato developmental processes. ### Competing Interest Statement The authors have declared no competing interest.
Future crops need to be sustainable in the face of climate change. Modern barley varieties have been bred for high productivity and quality; however, they have suffered considerable genetic erosion, losing crucial genetic diversity. This renders modern cultivars vulnerable to climate change and stressful environments. We highlight the potential to tailor crops to a specific environment by utilising diversity inherent in an adapted landrace population. Tapping into natural biodiversity, while incorporating information about local environmental and climatic conditions, allows targeting of key traits and genotypes, enabling crop production in marginal soils. We outline future directions for the utilisation of genetic resources maintained in landrace collections to support sustainable agriculture through germplasm development via the use of genomics technologies and big data.
Phytophthora is a genus of destructive plant pathogens. Certain species are damaging to native ecosystems, forestry, and the horticultural sector, and there is evidence of their dissemination in plant imports. Horticultural nurseries are central nodes of the plant trade and previous studies have found a high diversity of Phytophthora associated with plant nursery stock. It was subsequently hypothesized that green waste disposal sites in nurseries could harbour diverse Phytophthora communities and act as a pathogen reservoir and conduit, facilitating further Phytophthora infection of nursery stock and its spread into the wider environment. This project identified Phytophthora species associated with green waste at three Scottish nurseries by sampling material from waste piles, water run-off from piles, and roots from discarded plants. Species were identified using a baiting method and sequencing of environmental DNA. Plant nursery green waste was shown to harbour diverse and varied Phytophthora species assemblages, with differences among nurseries reflecting biosecurity management practices. Eighteen Phytophthora species were detected in the samples, including the highly destructive pathogens P. ramorum and P. austrocedri. Results suggest that the improved management of waste, for example through effective on-site composting, is essential to reduce the risk of Phytophthora pathogens spreading from nurseries into the wider environment.
This dataset on Zenodo accompanies the manuscript Schiffer-Forsyth et al. (2023), Phytophthora in horticultural nursery green waste – a risk to plant health. There are two files: metadata.tsv - plain text table as tab-separated variables raw_data.tar.gz - compressed archive of 81 paired raw FASTQ files This represents a complete Illumina MiSeq run, with the names of the unrelated samples redacted. To repeat the analysis described in the paper, first install THAPBI PICT. See https://github.com/peterjc/thapbi-pict/ for instructions. At the time of the paper, v0.14.1 was the current release (with a near-identical v1.0.0 expected to be released shortly). Next, decompress the raw data into a folder of paired gzipped FASTQ files. There is no need to decompress those: $ tar -zxvf raw_data.tar.gz $ ls -1 raw_data/If you wish, verify the checksums to confirm the data integrity: $ cd raw_data/ $ md5sum -c MD5SUM.txt $ cd ..Setup output directories: $ mkdir -p intermediate/ summary/You can run the analysis in one step. This should take under five minutes:$ thapbi_pict pipeline -i raw_data/ \ -n raw_data/SynCtrl_*.fastq.gz \ -y raw_data/SynCtrl_*.fastq.gz \ -s intermediate/ -o summary/ \ -t metadata.tsv -x 3 -c 1,2,4,5The options here are as follows: -i raw_data - input directory of paired raw FASTQ files. -n raw_data/SynCtrl_*.fastq.gz - negative controls used to increase the absolute abundance threshold -y raw_data/SynCtrl_*.fastq.gz - synthetic controls used to increase the fractional abundance threshold -s intermediate/ - optional location to store intermediate files -o summary/ - output location for reports -t metadata.tsv -x 3 -c 1,2,4,5 - show and sort on metadata columns 1, 2, 4 and 5 from metadata.tsv using column 3 to cross-reference the FASTQ filename stems (semi-colon separated lists for replicates). This assumes the following key default settings: -a 100 -f 0.001 - default absolute and fractional abundance thresholds -d - - default to the provided ITS1 database With these settings, only synthetic sequences were found in the controls, and therefore the thresholds were not automatically increased any further. Note some of these options could change in future releases of the software, and in particular there would likely be additional Phytophthora species or sequences in future updates to the default database. Output file summary/ITS1.samples.onebp.xlsx (and .tsv) is equivalent to Table 2 (after pooling replicates, and applying human judgement to resolve ambiguous ITS1 markers shared by multiple species). Note P. austrocedri was identified in three samples, N2-Water_S2 and N2-Water_S22 described in this work, and a third sample REDACTED_S28 from another location.
SUMMARYThe distribution of recombination events along large cereal chromosomes is uneven and is generally restricted to gene‐rich telomeric ends. To understand how the lack of recombination affects diversity in the large pericentromeric regions, we analysed deep exome capture data from a final panel of 815 Hordeum vulgare (barley) cultivars, landraces and wild barleys, sampled from across their eco‐geographical ranges. We defined and compared variant data across the pericentromeric and non‐pericentromeric regions, observing a clear partitioning of diversity both within and between chromosomes and germplasm groups. Dramatically reduced diversity was found in the pericentromeres of both cultivars and landraces when compared with wild barley. We observed a mixture of completely and partially differentiated single‐nucleotide polymorphisms (SNPs) between domesticated and wild gene pools, suggesting that domesticated gene pools were derived from multiple wild ancestors. Patterns of genome‐wide linkage disequilibrium, haplotype block size and number, and variant frequency within blocks showed clear contrasts among individual chromosomes and between cultivars and wild barleys. Although most cultivar chromosomes shared a single major pericentromeric haplotype, chromosome 7H clearly differentiated the two‐row and six‐row types associated with different geographical origins. Within the pericentromeric regions we identified 22 387 non‐synonymous SNPs, 92 of which were fixed for alternative alleles in cultivar versus wild accessions. Surprisingly, only 29 SNPs found exclusively in the cultivars were predicted to be ‘highly deleterious’. Overall, our data reveal an unconventional pericentromeric genetic landscape among distinct barley gene pools, with different evolutionary processes driving domestication and diversification.
The microbiota populating the rhizosphere, the interface between roots and soil, can modulate plant growth, development, and health. These microbial communities are not stochastically assembled from the surrounding soil, but their composition and putative function are controlled, at least partially, by the host plant. Here, we use the staple cereal barley as a model to gain novel insights into the impact of differential applications of nitrogen, a rate-limiting step for global crop production, on the host genetic control of the rhizosphere microbiota. Using a high-throughput amplicon sequencing survey, we determined that nitrogen availability for plant uptake is a factor promoting the selective enrichment of individual taxa in the rhizosphere of wild and domesticated barley genotypes. Shotgun sequencing and metagenome-assembled genomes revealed that this taxonomic diversification is mirrored by a functional specialization, manifested by the differential enrichment of multiple Gene Ontology terms, of the microbiota of plants exposed to nitrogen conditions limiting barley growth. Finally, a plant soil feedback experiment revealed that host control of the barley microbiota underpins the assembly of a phylogenetically diverse group of bacteria putatively required to sustain plant performance under nitrogen-limiting supplies. Taken together, our observations indicate that under nitrogen conditions limiting plant growth, host-microbe and microbe-microbe interactions fine-tune the host genetic selection of the barley microbiota at both taxonomic and functional levels. The disruption of these recruitment cues negatively impacts plant growth. IMPORTANCE The microbiota inhabiting the rhizosphere, the thin layer of soil surrounding plant roots, can promote the growth, development, and health of their host plants. Previous research indicated that differences in the genetic composition of the host plant coincide with variations in the composition of the rhizosphere microbiota. This is particularly evident when looking at the microbiota associated with input-demanding modern cultivated varieties and their wild relatives, which have evolved under marginal conditions. However, the functional significance of these differences remains to be fully elucidated. We investigated the rhizosphere microbiota of wild and cultivated genotypes of the global crop barley and determined that nutrient conditions limiting plant growth amplify the host control on microbes at the root-soil interface. This is reflected in a plant- and genotype-dependent functional specialization of the rhizosphere microbiota, which appears to be required for optimal plant growth. These findings provide novel insights into the significance of the rhizosphere microbiota for plant growth and sustainable agriculture.
The WHIRLY (WHY) DNA/RNA binding proteins fulfil multiple but poorly characterised functions in leaf development. Here, we show that WHY1 transcript levels were highest in the bases of 7-day old barley leaves. Immunogold labelling revealed that the WHY1 protein was more abundant in the nuclei than the proplastids of the leaf bases. To identify transcripts associated with leaf development we conducted hierarchical clustering of differentially abundant transcripts along the developmental gradient of wild-type leaves. Similarly, metabolite profiling was employed to identify metabolites exhibiting a developmental gradient. A comparative analysis of transcripts and metabolites in barley lines (W1–1 and W1–7) lacking WHY1, which show delayed greening compared with the wild type revealed that the transcript profile of leaf development was largely unchanged in W1–1 and W1–7 leaves. However, there were differences in levels of several transcripts encoding transcription factors associated with chloroplast development. These include a barley homologue of the Arabidopsis GATA transcription factor that regulates stomatal development, greening and chloroplast development, NAC1; two transcripts with similarity to Arabidopsis GLK1 and two transcripts encoding ARF transcriptions factors with functions in leaf morphogenesis and development. Chloroplast proteins were less abundant in the W1–1 and W1–7 leaves than the wild type. The levels of tricarboxylic acid cycle metabolites and GABA were significantly lower in WHY1 knockdown leaves than the wild type. This study provides evidence that WHY1 is localised in the nuclei of leaf bases, contributing the regulation of nuclear-encoded transcripts that regulate chloroplast development.
Moss-sedge heath, an important habitat of oceanic alpine regions, is under threat from elevated nitrogen (N) deposition and rising temperatures increasing decomposition rates and fragmentation of the extensive, thick moss mats of Racomitrium lanuginosum which characterise this habitat. Here, we examine the potential effects of N deposition and temperature on fungal communities associated with R. lanuginosum mats. Fungal community composition in both green moss shoots and underlying soil were significantly influenced by mat temperatures and green moss shoot N contents. Total OTU, Ascomycota, Basidiomycota and saprotrophic fungal richness in shoots were all positively related to temperature, while in soil, the richness of all groups, except Basidiomycota, was positively related to moss tissue N. The observed community changes suggest that increased N loading and increasing temperatures are ameliorating growing conditions for fungi associated with moss-sedge heath. Further study is required to determine if the observed changes in fungal communities contribute to accelerated decomposition of moss mats and degradation of the habitat.
A prerequisite to exploiting soil microbes for sustainable crop production is the identification of the plant genes shaping microbiota composition in the rhizosphere, the interface between roots and soil. Here, we use metagenomics information as an external quantitative phenotype to map the host genetic determinants of the rhizosphere microbiota in wild and domesticated genotypes of barley, the fourth most cultivated cereal globally. We identify a small number of loci with a major effect on the composition of rhizosphere communities. One of those, designated the QRMC-3HS , emerges as a major determinant of microbiota composition. We subject soil-grown sibling lines harbouring contrasting alleles at QRMC-3HS and hosting contrasting microbiotas to comparative root RNA-seq profiling. This allows us to identify three primary candidate genes, including a Nucleotide-Binding-Leucine-Rich-Repeat ( NLR ) gene in a region of structural variation of the barley genome. Our results provide insights into the footprint of crop improvement on the plant’s capacity of shaping rhizosphere microbes.
'Crumbly' fruit is a developmental disorder in raspberry that results in malformed and unsaleable fruits. For the first time, we define two distinct crumbly phenotypes as part of this work. A consistent crumbly fruit phenotype affecting the majority of fruits every season, which we refer to as crumbly fruit disorder (CFD) and a second phenotype where symptoms vary across seasons as malformed fruit disorder (MFD). Here, segregation of crumbly fruit of the MFD phenotype was examined in a full-sib family and three QTL (Quantitative Trait Loci) were identified on a high density GbS (Genotype by Sequencing) linkage map. This included a new QTL and more accurate location of two previously identified QTLs. A microarray experiment using normal and crumbly fruit at three different developmental stages identified several genes that were differentially expressed between the crumbly and non-crumbly phenotypes within the three QTL. Analysis of gene function highlighted the importance of processes that compromise ovule fertilization as triggers of crumbly fruit. These candidate genes provided insights regarding the molecular mechanisms involved in the genetic control of crumbly fruit in red raspberry. This study will contribute to new breeding strategies and diagnostics through the selection of molecular markers associated with the crumbly trait.
Species of Phytophthora, plant pathogenic eukaryotic microbes, can cause disease on many tree species. Genome sequencing of species from this genus has helped to determine components of their pathogenicity arsenal. Here, we sequenced genomes for two widely distributed species, Phytophthora pseudosyringae and Phytophthora boehmeriae, yielding genome assemblies of 49 and 40 Mb, respectively. We identified more than 270 candidate disease promoting RXLR effector coding genes for each species, and hundreds of genes encoding candidate plant cell wall degrading carbohydrate active enzymes (CAZymes). These data boost genome sequence representation across the Phytophthora genus, and form resources for further study of Phytophthora pathogenesis.