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Nicholson, P.

Publications and source records attributed to Nicholson, P..

At least 19 recordsLinked to original sources

Chromosome-level, haplotype-resolved genome assembly of the tanniferous forage legume big trefoil (Lotus pedunculatus Cav.) using CiFi

Big trefoil (Lotus pedunculatus Cav.) is a perennial forage legume that thrives on acidic, low-fertility soils and produces condensed tannins that reduce enteric methanogenesis in ruminants. Despite this agronomic potential, genomic resources for the species remain scarce, and the existing haploid assembly does not resolve the two haplotypes of this outcrossing diploid species. Here we present a haplotype-resolved, chromosome-level reference genome for L. pedunculatus genotype Lusitano29 -- the first plant genome assembled using CiFi, a long-read chromosome conformation capture method. We combined PacBio HiFi long reads with CiFi concatemers produced from DpnII and HindIII libraries; in silico digestion and combinatorial pairing of the resulting monomers yielded 790.3 M and 10.3 M pseudo-paired contacts, respectively, enabling scaffolding and manual curation to chromosome level. The 991.1 Mb assembly resolves two phased haplotypes of 500 and 491 Mb, with 96.6% of the sequence anchored in twelve pseudo-chromosomes (six per haplotype). Telomeric repeats were detected at 19 of 24 pseudo-chromosome ends, and no structural errors were detected (scaffold N50 73.8 Mb; consensus QV 64.7; k-mer completeness 99.4%; genome-mode BUSCO completeness 97.0%; CRAQ S-AQI 100.0). Annotation supported by PacBio Iso-Seq full-length transcripts predicted 38,069 and 36,484 protein-coding genes in haplotypes 1 and 2, respectively (protein-mode BUSCO completeness 96.5%), indicating a high completeness of annotated genes. This genome assembly provides a foundation for allele-aware trait dissection of proanthocyanidin biosynthesis, comparative genomics in Lotus, and population genomics and genomics-assisted breeding in L. pedunculatus.

genomics↗

A wheat immune receptor pair executes cell death through a helper MLKL

To promote disease resistance, plant nucleotide-binding, leucine-rich repeat (NLR) immune receptors often require paired co-receptors. In many cases, paired NLRs comprise one NLR to perceive effectors (the sensor) and another NLR to execute cell death (the helper). However, NLRs can also pair with sensor kinase fusion protein (KFP) receptors, but whether non-NLR components within such pairs can execute cell death, remains unclear. Here, we investigate the mechanism of an immune receptor pair comprising the wheat NLR Rwt3.6.8 NLR (R3NLR) and an MLKL protein, Rwt3.6.8 associated kinase (R3AK). Using Nicotiana benthamiana transient expression assays we confirmed that both R3NLR and R3AK are required for cell death in response to blast pathogen effectors PWT3, PWT6 or PWT8. Through mutational analysis we show the 4-helical bundle (4HB) domain of R3AK is required to execute cell death and R3AK can be made auto-active by perturbing the kinase catalytic active site. Activation of R3AK is also associated with a shift to a higher oligomeric state. Furthermore, as the NLR R3NLR is not actively involved in the execution of cell death we hypothesise that R3AK acts as a helper. A phylogenetic analysis indicates widespread distribution of this paired configuration in Poales. Together, this study establishes a novel resistance mechanism involving a non-canonical NLR/MLKL system. Significance StatementHere we investigate the mechanism of a novel plant immune receptor pair from wheat, R3NLR/R3AK. A nucleotide-binding, leucine-rich repeat (NLR) receptor and a mixed lineage kinase like (MLKL) protein are both required to mediate resistance to blast pathogen effector proteins PWT3, PWT6 and PWT8. Adopting a mutagenesis approach, we show that the MLKL protein executes cell death through its N-terminal 4-helical bundle domain, and this is associated with a shift to a higher oligomeric state. Mutations of conserved sequence motifs in the NLR support its role as a sensor, although effector interactions have not yet been observed. This study reveals a plant immune receptor pair that functions via a putative NLR sensor paired to a cell death executing MLKL protein.

plant biology↗

Chromosome-level genome assembly of the swallowtail butterfly Parides eurimedes mylotes is a valuable resource for studying wing coloration

High-quality chromosome-scale assemblies are scarce in Papilionidae. This limits comparative genomics to model species, Lepidoptera, Bombyx mori. Here, we present a phased, chromosome-level genome assembly of Parides eurimedes mylotes. We generated this assembly using 125[x] PacBio HiFi sequencing and assembled it with hifiasm. The final haplotype assemblies (Hap1 and Hap2) span 274 Mb and 270 Mb, respectively. These assemblies are organized into 31 near-telo-mere-to-telomere chromosomes, with scaffold N50 values of 9.72 Mb and 9.22 Mb, respectively. BUSCO analysis revealed assembly completeness of 96.6 % and 96.4 % for Hap1 and Hap2, respectively. Repeats annotation identified 18-19 % repetitive content, with Helitron elements being the dominant class of transposable elements. We identified the W and Z sex chromosomes and completely assembled the mitochondrial genome. Compared to the previously available Parides photinus draft assembly, our genome exhibits an 11,000-fold reduction in scaffold fragmentation and nearly complete gene assembly. This assembly provides a robust genomic reference for functional, evolutionary, and multi-omics investigation in Papilionidae. In addition to serving as a high-quality genomic reference for Papilionidae, this assembly is essential for linking the genetic architecture of butterfly wings to the hierarchical nanostructures underlying structural coloration. By identifying the genes and regulatory networks involved in scale morphogenesis, we can correlate the butterflys genotype with its photonic function. This insight into the evolutionary origin of the biological photonic systems informs the design of biomimetic, structurally colored materials.

genomics↗

Diverse Diterpenoid Phytoalexins Shape Wheat Chemical Defenses

Cereal crops rely on a wide array of specialized metabolites to defend themselves against microbial pathogens. Here, using a combination of genome-wide analysis, heterologous pathway reconstruction, structural elucidation and in planta validation, we identify two pathogen-induced diterpenoid pathways in wheat that produce diterpenoids: the new glycosylated diterpenes aspisoside A and aspisoside B, and the diterpene alcohols scutenol A and scutenol B. The pathogen-responsive nature of these pathways, together with their antimicrobial activity are consistent with a likely defensive role in wheat. These compounds are produced by two biosynthetic gene clusters, each encoding a discrete pathway, so revealing organizational separation of wheat diterpenoid-based chemical defenses. The aspisoside-producing gene cluster is syntenic with diterpenoid phytoalexin-producing clusters in rice and barley, for momilactone and hordedane biosynthesis, respectively, yet gives rise to structurally distinct phytoalexins in wheat. The scutenol cluster is syntenic with currently uncharacterized predicted biosynthetic gene clusters in barley, oat, and Brachypodium. These findings establish diterpene glycosides as a previously unrecognized component of wheat defense chemistry and provide new insights into the chemical diversification of defense-related biosynthetic gene clusters within the Poaceae.

plant biology↗

A segment of Triticum timopheevii chromosome 3G confers type II Fusarium head blight resistance and reduces DON accumulation in wheat.

Fusarium head blight (FHB) is a major disease of small grain cereals that is particularly damaging on wheat and can result in yield losses of up to 80%. Resistance to FHB in wheat is broadly classified as resistance to initial infection (type I) or resistance to disease spread within the spike (type II). A high level of type I FHB resistance was reported in an accession of wheat wild relative Triticum timopheevii. Hexaploid wheat-T. timopheevii introgression lines carrying a segment of the short arm of chromosome 3G (Chr3G) of this accession expressed high levels of FHB resistance following spray inoculation. Further analyses of these introgression lines showed that the Chr3G segment confers a potent type II resistance, accompanied by improved grain weight retention, and reduced deoxynivalenol (DON) accumulation in grain. These results indicate that the Chr3G resistance has the potential to dramatically reduce FHB susceptibility and DON accumulation in grain in wheat cultivars. An introgression of a segment of Chr7At into the short arm of chromosome Chr7A also enhanced type II FHB resistance.

plant biology↗

Chromosome-level phased genome assembly of the Argan tree Sideroxylon spinosum L.

Argan (Sideroxylon spinosum L. previously known as Argania spinosa) is an endemic tree of Morocco, primarily used for seed oil extraction. Increasing interest in its biology and in genes relevant for oil quality and stress resistance demands for high-quality models of its genome and transcriptome. We integrated sequencing data from PacBio HiFi long-reads and Illumina Hi-C to generate independently assembled phased genome models for both parental haplotigs with lengths of 636Mb and 655 Mb, respectively, and with a BUSCO completeness score of >97.8%. Both haplotigs consist of 11 independently assembled fully resolved telomere-to-telomere chromosomes, similar to other species of the Sapotaceae (n = 10-13), and they both contain approximately [~]60% of highly repetitive regions. Comparative analysis with other Sapotaceous genomes suggests overall chromosome conservation, with repeat expansion and chromosome fusion for the two largest chromosomes (chr1, chr2). We also independently assembled the chloroplast genome. This high-quality genome assembly provides a valuable resource for advancing future research on argan biology, its genetic diversity, and traits relevant for fitness and oil production.

genomics↗

The wheat powdery mildew resistance gene Pm4 also confers resistance to wheat blast

Wheat blast, caused by the fungus Magnaporthe oryzae, threatens global cereal production since its emergence in Brazil in 1985 and recently spread to Bangladesh and Zambia. Here we demonstrate that the AVR-Rmg8 effector, common in wheat-infecting isolates, is recognised by the gene Pm4, previously shown to confer resistance to specific races of Blumeria graminis f.sp. tritici, the cause of powdery mildew of wheat. We show that Pm4 alleles differ in their recognition of different AVR-Rmg8 alleles, and some confer resistance only in seedling leaves but not spikes making it important to select for those alleles that function in both tissues. This study has identified a gene recognising an important virulence factor present in wheat blast isolates in Bangladesh and Zambia and represents an important first step towards developing durably resistant wheat cultivars for these regions.

plant biology↗

Barley cultivars shape the abundance, phenotype, genotype and gene expression of their associated microbiota by differential root exudate secretion

Plant associated microbes play vital roles in promoting plant growth and health, with plants secreting root exudates into the rhizosphere to attract beneficial microbes. Exudate composition defines the nature of microbial recruitment, with different plant species attracting distinct microbiota to enable optimal adaptation to the soil environment. To more closely examine the relationship between plant genotype and microbial recruitment, we analysed the rhizosphere microbiomes of landrace (Chevallier) and modern (NFC Tipple) barley cultivars. Distinct differences were observed between the plant-associated microbiomes of the two cultivars, with the plant-growth promoting rhizobacterial genus Pseudomonas substantially more abundant in the Tipple rhizosphere. Striking differences were also observed between the phenotypes of recruited Pseudomonas populations, alongside distinct genotypic clustering by cultivar. Cultivar-driven Pseudomonas selection was driven by root exudate composition, with the greater abundance of hexose sugars secreted from Tipple roots attracting microbes better adapted to growth on these metabolites, and vice versa. Cultivar-driven selection also operates at the molecular level, with both gene expression and the abundance of ecologically relevant loci differing between Tipple and Chevallier Pseudomonas isolates. Finally, cultivar-driven selection is important for plant health, with both cultivars showing a distinct preference for microbes selected by their genetic siblings in rhizosphere transplantation assays.

microbiology↗

Single-cell characterisation of the hematopoietic bone marrow interactome in health and disease

1The bone marrow (BM) is a complex microenvironment and the primary site of hematopoiesis, coordinating the production of billions of blood cells every day. Despite the essential role of the hematopoietic niche in maintaining hemostasis and its relevance to hematopoietic diseases, many aspects of this environment remain poorly characterised due to experimental hurdles. Here we present a high-resolution characterisation of the niche in health and acute myeloid leukemia (AML) by establishing a comprehensive single-cell gene expression database of nearly 340,000 BM constituent cells encompassing all disease stages (healthy BM, AML at diagnosis, remission and relapse). We characterised the cell type composition of the BM and found that the proportions of both myeloid and lymphoid lineage cell types are significantly altered in AML. We also determined broadscale dysregulation of gene expression in almost all BM cell types upon establishment of AML, indicating that the entire niche is disrupted by the disease. Given the importance of interactions between hematopoietic cells and their microenvironment in regulating their function and properties, we determined all possible ligand-receptor interactions between hematopoietic stem and progenitor cells (HSPC) and every other BM constituent cell type. This analysis revealed a remarkable expansion of HSPC interactions in AML involving multiple BM constituent cells that can drive dysregulated HSPC-cell adhesion, immunosuppression and enhanced cytokine signalling. In particular, we found that interactions involving TGFB1 become widespread in AML and present evidence that these interactions can drive AML cell quiescence in vitro, thus highlighting TGFB1 signalling as a potential target for increasing drug sensitivity and preventing relapse. Our results shed light on potential mechanisms of enhanced competitiveness of AML HSPCs and an overall skewed microenvironment that fosters AML growth.

cancer biology↗

Identification of Fusarium Head Blight resistance in Triticum timopheevii accessions and characterisation of wheat-T. timopheevii introgression lines for enhanced resistance.

A diverse panel of wheat wild relative species was screened for resistance to Fusarium head blight (FHB) by spray inoculation. The great majority of species and accessions were susceptible or highly susceptible to FHB. Accessions of Triticum timopheevii (P95-99.1-1), Agropyron desertorum (9439957) and Elymus vaillantianus (531552) were highly resistant to FHB while additional accessions of T. timopheevii were found to be susceptible to FHB. A combination of spray and point inoculation assessments over two consecutive seasons indicated that the resistance in accession P95-99.1-1 was due to enhanced resistance to initial infection of the fungus (type 1 resistance), and not to reduction in spread (type 2 resistance). A panel of wheat-T. timopheevii (accession P95-99.1-1) introgression lines was screened for FHB resistance over two consecutive seasons using spray inoculation. Most introgression lines were similar in susceptibility to FHB as the wheat recipient (Paragon) but substitution of the terminal portion of chromosome 3BS of wheat with a similar-sized portion of 3G of T. timopheevii significantly enhanced FHB resistance in the wheat background.

plant biology↗

Tissue-Specific Transcriptome Responses to Fusarium Head Blight and Fusarium Root Rot

Fusarium head blight (FHB) and Fusarium root rot (FRR) are important diseases of small-grained cereals caused by Fusarium species. While host response to FHB has been subject to extensive study, very little is known about response to FRR and the transcriptome responses of FHB and FRR have not been thoroughly compared. Brachypodium distachyon (Bd) is an effective model for investigating host responses to both FHB and FRR. In this study the transcriptome response of Bd to F. graminearum (Fg) infection of heads and roots was investigated. An RNA-seq analysis was performed on both Bd FHB and FRR during the early infection. Additionally, an RNA-seq analysis was performed on in vitro samples of Fg for comparison with Fg gene expression in planta. Differential gene expression and gene-list enrichment analyses were used to compare FHB and FRR transcriptome responses in both Bd and Fg. Differential expression of selected genes was confirmed using RT-qPCR. Most genes associated with receptor signalling, cell-wall modification, oxidative stress metabolism, and cytokinin and auxin biosynthesis and signalling genes were generally upregulated in FHB or were downregulated in FRR. In contrast, Bd genes involved in jasmonic acid and ethylene biosynthesis and signalling, and antimicrobial production were similarly differentially expressed in both tissues in response to infection. A transcriptome analysis of predicted Fg effectors with the same infected material revealed elevated expression of both core tissue- independent genes including cell-wall degradation enzymes and the gene cluster for DON production but also several tissue-dependent genes including those for aurofusarin production and cutin degradation. This evidence suggests that Fg modulates its transcriptome to different tissues of the same host.

plant biology↗

Molecular characterization of cell types in the squid Loligo vulgaris

Cephalopods have long been getting a lot of attention for their fascinating behavioral abilities and for the complexity of their nervous systems that set them apart from other mollusks. Because of the great evolutionary distance that separates vertebrates from mollusks, it is evident that higher cognitive features have evolved independently in this clade although they sometimes resemble cognitive functions of vertebrates. Alongside their complex behavioral abilities, cephalopods have evolved specialized cells and tissues, such as the chromatophores for camouflage or suckers to grasp prey. Gaining a better understanding of the biology of various species of cephalopods can significantly improve our knowledge of how these animals evolved and better identify the mechanisms that drive the astonishing faculties of their nervous systems. In this study, we performed single-cell transcriptomics of whole heads of Loligo vulgaris pre-hatchlings. We characterized the different cell types in the head of these animals and explored the expression patterns of core cell type markers by hybridization chain reaction. We were able to thoroughly describe some major components of the squid nervous that play important roles for the maintenance, development and sensory function in the nervous system of these animals.

developmental biology↗

A wheat kinase and immune receptor form the host-specificity barrier against the blast fungus

Since emerging in Brazil in 1985, wheat blast has spread throughout South America and recently appeared in Bangladesh and Zambia. We show that two wheat resistance genes, Rwt3 and Rwt4, acting as host-specificity barriers against non-Triticum blast pathotypes encode a nucleotide-binding leucine-rich repeat immune receptor and a tandem kinase, respectively. Molecular isolation of these genes allowed us to develop assays that will ensure the inclusion of these two genes in the wheat cultivars to forestall the recurrence of blast host jumps.

plant biology↗

Genome Mining Uncovers Clustered Biosynthetic Pathways for Defense-Related Molecules in Bread Wheat

Wheat is one of the most widely grown food crops in the world. However, it succumbs to numerous pests and pathogens that cause substantial yield losses. A better understanding of biotic stress responses in wheat is thus of major importance. Here we identify previously unknown pathogen-induced biosynthetic pathways that produce a diverse set of molecules, including flavonoids, diterpenes and triterpenes. These pathways are encoded by six biosynthetic gene clusters and share a common regulatory network. We further identify associations with known or novel phytoalexin clusters in other cereals and grasses. Our results significantly advance the understanding of chemical defenses in wheat and open up new avenues for enhancing disease resistance in this agriculturally important crop.

plant biology↗

Developmental changes and metabolic reprogramming during establishment of infection and progression of Trypanosoma brucei brucei through its insect host

Trypanosoma brucei ssp., unicellular parasites causing human and animal trypanosomiasis, are transmitted between mammals by tsetse flies. Periodic changes in variant surface glycoproteins (VSG), which form the parasite coat in the mammal, allow them to evade the host immune response. Different isolates of T. brucei show heterogeneity in their repertoires of VSG genes and have single nucleotide polymorphisms and indels that can impact on genome editing. T. brucei brucei EATRO1125 (AnTaR1 serodeme) is an isolate that is used increasingly often because it is pleomorphic in mammals and fly transmissible, two characteristics that have been lost by the most commonly used laboratory stocks. We present a genome assembly of EATRO1125, including contigs for the intermediate and mini-chromosomes that serve as repositories of VSG genes. In addition, de novo transcriptome assemblies were performed using Illumina sequences from tsetse-derived trypanosomes. Reads of 150 bases enabled closely related members of multigene families to be discriminated. This revealed that the transcriptome of midgut-derived parasites is dynamic, starting with the expression of high affinity hexose transporters and glycolytic enzymes and then switching to proline uptake and catabolism. These changes resemble the transition from early to late procyclic forms in culture. Further metabolic reprogramming, including upregulation of tricarboxylic acid cycle enzymes, occurs in the proventriculus. Many transcripts upregulated in the salivary glands encode surface proteins, among them 7 metacyclic VSGs, multiple BARPs and GCS1/HAP2, a marker for gametes. A novel family of transmembrane proteins, containing polythreonine stretches that are predicted to be O-glycosylation sites, was also identified. Finally, RNA-Seq data were used to create an optimised annotation file with 5 and 3 untranslated regions accurately mapped for 9302 genes. We anticipate that this will be of use in identifying transcripts obtained by single cell sequencing technologies. AUTHOR SUMMARYTrypanosoma brucei ssp. are single-celled parasites that cause two tropical diseases: sleeping sickness in humans and nagana in domestic animals. Parasites survive in the host bloodstream because they periodically change their surface coats and also because they can switch from slender dividing forms to stumpy non-dividing forms. The latter can be transmitted to their second host, the tsetse fly. Although closely related, different geographical isolates differ in their repertoire of surface coats and have small, but important differences in their DNA sequences. In addition, laboratory strains that are transferred between mammals by needle passage lose the ability to produce stumpy forms and to infect flies. The isolate T. b. brucei EATRO1125 is often used for research as it produces stumpy forms and is fly transmissible. We provide an assembly of the genome of this isolate, including part of the repertoire of coat proteins, and a detailed analysis of the genes that the parasites express as they establish infection and progress through the fly. This has provided new insights into trypanosome biology. The combined genomic (DNA) and transcriptomic (RNA) data will be useful resources for the trypanosome research community.

microbiology↗

Evolution of the bread wheat D-subgenome and enriching it with diversity from Aegilops tauschii

Aegilops tauschii, the diploid wild progenitor of the D-subgenome of bread wheat, constitutes a reservoir of genetic diversity for improving bread wheat performance and environmental resilience. To better define and understand this diversity, we sequenced 242 Ae. tauschii accessions and compared them to the wheat D-subgenome. We characterized a rare, geographically-restricted lineage of Ae. tauschii and discovered that it contributed to the wheat D-subgenome, thereby elucidating the origin of bread wheat from at least two independent hybridizations. We then used k-mer-based association mapping to identify discrete genomic regions with candidate genes for disease and pest resistance and demonstrated their functional transfer into wheat by transgenesis and wide crossing, including the generation of a library of synthetic hexaploids incorporating diverse Ae. tauschii genomes. This pipeline permits rapid trait discovery in the diploid ancestor through to functional genetic validation in a hexaploid background amenable to breeding.

plant biology↗

Different effects of phytohormones on Fusarium head blight and Fusarium root rot resistance in Brachypodium distachyon

Fusarium graminearum is a devastating pathogen of small grain cereals causing both Fusarium head blight (FHB) and Fusarium root rot (FRR). Exogenous application of phytohormones has been shown to affect FHB resistance. In contrast to FHB, FRR remains poorly characterised and it is unknown whether phytohormones play similar roles in FHB and FRR. In this present study, B. distachyon floral tissues at mid-anthesis and root tissues from seedlings were exogenously treated with several phytohormones before inoculation with F. graminearum. The canonical defence-associated phytohormones had differing effects on FHB and FRR. Salicylic acid (SA) significantly increased susceptibility to FRR but not to FHB while jasmonic acid (JA) and ethylene increased resistance to FRR but increased susceptibility to FHB. Additionally, the growth-associated phytohormones auxin and cytokinin significantly increased resistance and susceptibility, respectively, to both diseases. This study is the first to compare phytohormone effects between FHB and FRR in the same host. HighlightThe tissue-dependent effects of defence phytohormones and tissue-independent effects of development phytohormones on F. graminearum-induced Fusarium head blight and Fusarium root rot diseases in the model cereal Brachypodium distachyon.

plant biology↗

Type II Fusarium head blight susceptibility factor identified in wheat

Fusarium head blight (FHB) causes significant grain yield and quality reductions in wheat and barley. Most wheat varieties are incapable of preventing FHB spread through the rachis, but disease is typically limited to individually infected spikelets in barley. We point inoculated wheat lines possessing barley chromosome introgressions to test whether FHB resistance could be observed in a wheat genetic background. The most striking differential was between 4H(4D) substitution and 4H addition lines. The 4H addition line was similarly susceptible to the wheat parent, but the 4H(4D) substitution line was highly resistant, which suggests that there is an FHB susceptibility factor on wheat chromosome 4D. Point inoculation of Chinese Spring 4D ditelosomic lines demonstrated that removing 4DS results in high FHB resistance. We genotyped four Chinese Spring 4DS terminal deletion lines to better characterise the deletions in each line. FHB phenotyping indicated that lines del4DS-2 and del4DS-4, containing smaller deletions, were susceptible and had retained the susceptibility factor. Lines del4DS-3 and del4DS-1 contain larger deletions and were both significantly more resistant, and hence had presumably lost the susceptibility factor. Combining the genotyping and phenotyping results allowed us to refine the susceptibility factor to a 31.7 Mbp interval on 4DS. HighlightWe have identified a Type II Fusarium head blight susceptibility factor on the short arm of wheat chromosome 4D and refined its position to a 31.7 Mbp interval.

genetics↗