Arable-ley rotations can alleviate soil degradation and erosion. ● Multispecies leys can improve livestock health and reduce greenhouse gas emissions. ● Ley botanical composition is crucial for determining benefits. ● Lack of livestock infrastructure in arable areas may prevent arable-ley uptake. ● Long-term (10–25 years) research is needed to facilitate evidence-based decisions. Agricultural intensification and the subsequent decline of mixed farming systems has led to an increase in continuous cropping with only a few fallow or break years, undermining global soil health. Arable-ley rotations incorporating temporary pastures (leys) lasting 1–4 years may alleviate soil degradation by building soil fertility and improving soil structure. However, the majority of previous research on arable-ley rotations has utilized either grass or grass-clover leys within ungrazed systems. Multispecies leys, containing a mix of grasses, legumes, and herbs, are rapidly gaining popularity due to their promotion in agri-environment schemes and potential to deliver greater ecosystem services than conventional grass or grass-clover leys. Livestock grazing in arable-ley rotations may increase the economic resilience of these systems, despite limited research of the effects of multispecies leys on ruminant health and greenhouse gas emissions. This review aims to evaluate previous research on multispecies leys, highlighting areas for future research and the potential benefits and disbenefits on soil quality and livestock productivity. The botanical composition of multispecies leys is crucial, as legumes, deep rooted perennial plants (e.g., Onobrychis viciifolia and Cichorium intybus) and herbs (e.g., Plantago lanceolata) can increase soil carbon, improve soil structure, reduce nitrogen fertilizer requirements, and promote the recovery of soil fauna (e.g., earthworms) in degraded arable soils while delivering additional environmental benefits (e.g., biological nitrification inhibition and enteric methane reduction). Multispecies leys have the potential to deliver biologically driven regenerative agriculture, but more long-term research is needed to underpin evidence-based policy and farmer guidance.

Computerized treatments have been shown to be effective in young people with anxiety disorders. However, there has been limited investigation into the experiences of adolescents in undertaking this treatment. This qualitative study explored adolescents' experiences of being treated for an anxiety disorder, using an online intervention with therapist support, as part of a randomised controlled trial. Individual semi-structured interviews were conducted with thirteen adolescents aged between 13 and 17 years, who had received the online intervention. Data were analysed using reflexive thematic analysis. Two cross-cutting themes were identified: (i) Usability of the program, and (ii) Putting techniques into practice. Although the convenience of online sessions was recognised as positive, many of the adolescents expressed a preference for face-to-face sessions. The length of sessions and the large amount of reading involved was highlighted by most participants. The transdiagnostic nature of the content meant some elements of the programme were seen as less relevant. While many participants appreciated the 'fun' aspect of the program, others felt that certain aspects of the program were not suitable for their age group. Exposure, psychoeducation and cognitive restructuring were generally received positively, whereas responses to the use of relaxation and rewards were much more mixed. Our findings highlight the potential utility for online treatments delivered with therapist support. However, they also highlight how issues with both the format and treatment components can impact the acceptability of a program. As such, further work is warranted to improve the acceptability of online treatments for adolescents with anxiety disorders.

PURPOSE:This study tests the value of a measure of eating disorder (ED) psychopathology in predicting outcome following guided self-help in a non-underweight sample with regular binge eating. It examines whether early reductions in ED psychopathology are associated with remission status at post-treatment.METHODS:Seventy-two adults with bulimia nervosa, binge-eating disorder, or an atypical form of these illnesses received up to ten sessions of cognitive behaviour therapy-based guided self-help. Using a session-by-session measure of eating pathology and associated reliable change indices, response was analysed using receiver operating characteristic analysis to predict outcomes at post-treatment.RESULTS:In this routine care setting, nearly one-quarter of the sample achieved remission following GSH, approximately two-thirds of whom showed early change in ED psychopathology. Early change prior to session 6 was accurate in predicting later remission. Individuals showing early change did not differ from others on baseline characteristics or rates of attrition.CONCLUSION:Data suggest that a majority of those who respond to treatment will do so before the second half of treatment, information that could be used to ensure that evidence-based treatments are used as effectively as possible.LEVEL OF EVIDENCE:Level III.

Background & Aim Adoptive immunotherapy with Epstein-Barr virus (EBV)-specific T cells (VST) has proven effective for patients with refractory EBV-related malignancies. Two principal manufacturing approaches have been utilized to generate sufficient allogeneic T cells for therapy; sequential stimulation of donor PBMC with autologous EBV-positive lymphoblastic cell lines (LCLs), or in vitro expansion of interferon-gamma selected EBV-VST identified using EBV peptides. The Scottish National Blood Transfusion Service has a LCL-based EBV-VST bank which has been used to treat refractory PTLD patients for the last ten years on a Specials basis. This bank is being replaced by a next-generation bank based on EBV peptide selection and expansion in GMP-compliant closed-process manufacture. We have investigated the therapeutic material from both manufacturing processes and compared the phenotypic and functional characteristics of both to determine comparability. Methods, Results & Conclusion VST from both manufacturing processes were assessed for surface and intracellular markers by flow cytometry, which indicated that there were no significant differences in most parameters including CD3 content, CD4:CD8 ratio, and differentiation status. Peptide-generated products did demonstrate a significantly increased central memory compartment and mean IFNg expression. Further transcriptional analysis was undertaken using a custom TaqMAN low density array for relevant chemokine receptors, and indicated that EBV peptide-isolated VST had significantly increased levels of CCR7, CXCR4 and CXCR5, which correlate strongly with enhanced expansion and retention of central memory T cells. Transcriptional and phenotypic results were correlated with functional cytokine and migration assay to confirm responses. LCL-generated VST products showed significantly increased levels of CD38, indicating an accumulation of terminally differentiated functional T cells with reduced proliferative potential. The results from this study indicate that both EBV-VST manufacturing methods generate effective functional cells, but peptide-based VST demonstrate enhanced central memory cell retention which may predicate towards improved persistence in PTLD recipients. Adoptive immunotherapy with Epstein-Barr virus (EBV)-specific T cells (VST) has proven effective for patients with refractory EBV-related malignancies. Two principal manufacturing approaches have been utilized to generate sufficient allogeneic T cells for therapy; sequential stimulation of donor PBMC with autologous EBV-positive lymphoblastic cell lines (LCLs), or in vitro expansion of interferon-gamma selected EBV-VST identified using EBV peptides. The Scottish National Blood Transfusion Service has a LCL-based EBV-VST bank which has been used to treat refractory PTLD patients for the last ten years on a Specials basis. This bank is being replaced by a next-generation bank based on EBV peptide selection and expansion in GMP-compliant closed-process manufacture. We have investigated the therapeutic material from both manufacturing processes and compared the phenotypic and functional characteristics of both to determine comparability. VST from both manufacturing processes were assessed for surface and intracellular markers by flow cytometry, which indicated that there were no significant differences in most parameters including CD3 content, CD4:CD8 ratio, and differentiation status. Peptide-generated products did demonstrate a significantly increased central memory compartment and mean IFNg expression. Further transcriptional analysis was undertaken using a custom TaqMAN low density array for relevant chemokine receptors, and indicated that EBV peptide-isolated VST had significantly increased levels of CCR7, CXCR4 and CXCR5, which correlate strongly with enhanced expansion and retention of central memory T cells. Transcriptional and phenotypic results were correlated with functional cytokine and migration assay to confirm responses. LCL-generated VST products showed significantly increased levels of CD38, indicating an accumulation of terminally differentiated functional T cells with reduced proliferative potential. The results from this study indicate that both EBV-VST manufacturing methods generate effective functional cells, but peptide-based VST demonstrate enhanced central memory cell retention which may predicate towards improved persistence in PTLD recipients.

Root research on field grown crops is hindered by the difficulty of estimating root biomass in soil. Root washing, the current standard method is laborious and expensive. Biochemical methods to quantify root biomass in soil, targeting species-specific DNA, have potential as a more efficient assay. We combined an efficient DNA extraction method, designed specifically to extract DNA from soil, with well-established quantitative PCR methods to estimate the root biomass of twenty-two wheat varieties grown in field trials over two seasons. We also developed an assay for estimating root biomass for black-grass, a common weed of wheat cultivation. Two robust qPCR assays were developed to estimate the quantity of plant root DNA in soil samples, one specific to wheat and barley, and a second specific to black-grass. The DNA qPCR method was comparable, with high correlations, with the results of root washing from soil cores taken from winter wheat field trials. The DNA qPCR assay showed both variety and depth as significant factors in the distribution of root biomass in replicated field trials. The results suggest that these DNA qPCR assays are a useful, high throughput tool for investigating the genetic basis of wheat root biomass distribution in field grown crops, and the impact of black-grass root systems on crop production.

Abstract Onobrychis viciifolia (sainfoin) is a forage legume crop with many positive agronomic, environmental, nutritional and nutraceutical attributes. Farmers also benefit from its drought tolerance in areas of low rainfall and light free draining soil, mainly due to its deep taproot. It is resistant to most common pest and diseases and is a valuable resource for pollinators, specifically cultivated for honey production in some regions. It has many benefits for animals, being highly palatable and without danger of bloat, which can be life-threatening to livestock. Its decline in Northern Europe started during the Green Revolution and was impacted by changes towards more intensive farming. Unlike other forage legume crops such as red clover and lucerne, sainfoin does not respond well to inputs and is difficult to establish and maintain. Sainfoin could be classified as an ‘orphan crop’ with very little genetic improvement or agronomic studies in the past 60 years. In the past 5–10 years, however, there has been a resurgence in interest and this has given rise to a number of studies and initiation of systematic improvement of the crop, which is indispensable to its reintroduction into the farmed environment. Interest has been driven in part by considerable evidence to suggest that condensed tannins present in the legume foliage, together with other polyphenol compounds, have positive effects on animal nutrition together with anthelmintic properties. These compounds are also thought to play a role in environmental benefits. There remain many challenges to address in order to optimize the potential for cultivation of sainfoin and its use as a beneficial forage crop. This review makes particular reference to a recently completed project; ‘Legume Plus’, funded by the European Union and embracing a multi-disciplinary approach to both understand and improve the crop for farmers. The present review covers results from both this project and other studies during the past 5 years, also drawing on historic studies of etymology, taxonomy, genetics, agronomy and botany, aiming to be a useful resource for research and for practical plant breeders and agronomists.

A procedure based on 13C CPMAS NMR was developed to study procyanidins (PCs) and prodelphinidins (PDs) directly in milled sainfoin plant tissues. Blackcurrant and Tilia samples enabled reference spectra of purified proanthocyanidin (PA) fractions, crude extracts, and milled plant tissues, with characteristic resonances at 155, 144, and 132 ppm. PC/PD ratios were estimated from the I132/I155 intensity ratio and differed by 2.5 to 5.9% compared to thiolysis data. Normalization to the 155 ppm signal intensity from reference spectra enabled analysis of PA contents with an error of ca. 8 g PAs/100 g plant tissue. The procedure estimates the lignin contribution and allows for a correction of the PA content. In six sainfoin accessions, estimated PA contents were 1.6- to 20.8-fold higher than the thiolysis and 1.4- to 2.6-fold higher than the HCl-butanol-acetone results. Method differences may reflect the presence of unextractable, possibly high molecular weight PAs in sainfoin.

In the last decade, computational processing of big datasets has facilitated the analyses of unprecedented quantities of biological data. Thus, automation and big data analysis have been revolutionary in detecting and quantifying subtle phenotypes in cell biological contexts. Analyzing similar quantities of data in larger and more complicated biological systems such as developing embryos has been more challenging due to experimental limitations on both ensemble data collection and analysis. These challenges include photosensitivity of living samples and of the fluorescently tagged proteins under study, collectively limiting the number of images that can be acquired in a single timelapse series. Here we present a streamlined workflow to quantify dynamics of fluorescently labeled proteins over the course of several cell cycles in early embryos, taking advantage of the stereotypical nature of early development that is inherent for many organisms. We benchmark this pipeline studying a fluorescently labeled histone during early embryonic development of the nematode Caenorhabditis elegans. Our strategy allowed us to overcome biological and experimental variation among our timelapse series and quantify nuclear accumulation rate, chromatin incorporation, and turnover/stability of canonical histones. We find that histone proteins are broadly stable in early C. elegans development. Thus, changes in genome regulation occurring in early development do not manifest in gross changes in histone metabolism. Our method enabling characterization of cumulative protein dynamics over several cell cycles of developmental time with high temporal resolution can be applied to expand our understanding of diverse cellular and developmental processes.

Condensed tannins (CTs) account for up to 20% of the dry matter in forage legumes used as ruminant feeds. Beneficial animal responses to CTs have included improved growth, milk and wool production, fertility, and reduced methane emissions and ammonia volatilization from dung or urine. Most important is the ability of such forages to combat the effects of gastrointestinal parasitic nematodes. Inconsistent animal responses to CTs were initially attributed to concentration in the diet, but recent research has highlighted the importance of their molecular structures, as well as concentration, and also the composition of the diet containing the CTs. The importance of CT structural traits cannot be underestimated. Interdisciplinary research is the key to unraveling the relationships between CT traits and bioactivities and will enable future on-farm exploitation of these natural plant compounds. Research is also needed to provide plant breeders with guidelines and screening tools to optimize CT traits, in both the forage and the whole diet. In addition, improvements are needed in the competitiveness and agronomic traits of CT-containing legumes and our understanding of options for their inclusion in ruminant diets. Farmers need varieties that are competitive in mixed swards and have predictable bioactivities. This review covers recent results from multidisciplinary research on sainfoin (Onobrychis Mill. spp.) and provides an overview of current developments with several other tanniniferous forages. Tannin chemistry is now being linked with agronomy, plant breeding, animal nutrition, and parasitology. The past decade has yielded considerable progress but also generated more questions-an enviable consequence of new knowledge!

The size of mitotic chromosomes is coordinated with cell size in a manner dependent on nuclear trafficking. In this study, we conducted an RNA interference screen of the Caenorhabditis elegans nucleome in a strain carrying an exceptionally long chromosome and identified the centromere-specific histone H3 variant CENP-A and the DNA decatenizing enzyme topoisomerase-II (topo-II) as candidate modulators of chromosome size. In the holocentric organism C. elegans, CENP-A is positioned periodically along the entire length of chromosomes, and in mitosis, these genomic regions come together linearly to form the base of kinetochores. We show that CENP-A protein levels decreased through development coinciding with chromosome-size scaling. Partial loss of CENP-A protein resulted in shorter mitotic chromosomes, consistent with a role in setting chromosome length. Conversely, topo-II levels were unchanged through early development, and partial topo-II depletion led to longer chromosomes. Topo-II localized to the perimeter of mitotic chromosomes, excluded from the centromere regions, and depletion of topo-II did not change CENP-A levels. We propose that self-assembly of centromeric chromatin into an extended linear array promotes elongation of the chromosome, whereas topo-II promotes chromosome-length shortening.

A minority of children and adolescents with mental health problems access treatment. The reasons for poor rates of treatment access are not well understood. As parents are a key gatekeeper to treatment access, it is important to establish parents’ views of barriers/facilitators to accessing treatment. The aims of this study are to synthesise findings from qualitative and quantitative studies that report parents’ perceptions of barriers/facilitators to accessing treatment for mental health problems in children/adolescents. A systematic review and narrative synthesis were conducted. Forty-four studies were included in the review and were assessed in detail. Parental perceived barriers/facilitators relating to (1) systemic/structural issues; (2) views and attitudes towards services and treatment; (3) knowledge and understanding of mental health problems and the help-seeking process; and (4) family circumstances were identified. Findings highlight avenues for improving access to child mental health services, including increased provision that is free to service users and flexible to their needs, with opportunities to develop trusting, supportive relationships with professionals. Furthermore, interventions are required to improve parents’ identification of mental health problems, reduce stigma for parents, and increase awareness of how to access services.

Centromeres epitomize the central problem of propagating non-DNA sequence-based information across generations. In this issue of Developmental Cell, Hori et al. (2017) and French et al. (2017) show that the centromere-associated protein KNL-2/M18BP1 reads the centromere epigenetic code to maintain centromere identity.

For centuries, light microscopy has been a key method in biological research, from the early work of Robert Hooke describing biological organisms as cells, to the latest in live-cell and single-molecule systems. Here, we introduce some of the key concepts related to the development and implementation of modern microscopy techniques. We briefly discuss the basics of optics in the microscope, super-resolution imaging, quantitative image analysis, live-cell imaging, and provide an outlook on active research areas pertaining to light microscopy.

BACKGROUND:Sainfoin is a perennial forage legume with beneficial properties for animal husbandry due to the presence of secondary metabolites. However, worldwide cultivation of sainfoin is marginal due to the lack of varieties with good agronomic performance, adapted to a broad range of environmental conditions. Little is known about the genetics of sainfoin and only few genetic markers are available to assist breeding and genetic investigations. The objective of this study was to develop a set of SSR markers useful for genetic studies in sainfoin and their characterization in diverse germplasm.RESULTS:A set of 400 SSR primer combinations were tested for amplification and their ability to detect polymorphisms in a set of 32 sainfoin individuals, representing distinct varieties or landraces. Alleles were scored for presence or absence and polymorphism information content of each SSR locus was calculated with an adapted formula taking into account the tetraploid character of sainfoin. Relationships among individuals were visualized using cluster and principle components analysis. Of the 400 primer combinations tested, 101 reliably detected polymorphisms among the 32 sainfoin individuals. Among the 1154 alleles amplified 250 private alleles were observed. The number of alleles per locus ranged from 2 to 24 with an average of 11.4 alleles. The average polymorphism information content reached values of 0.14 to 0.36. The clustering of the 32 individuals suggested a separation into two groups depending on the origin of the accessions.CONCLUSIONS:The SSR markers characterized and tested in this study provide a valuable tool to detect polymorphisms in sainfoin for future genetic studies and breeding programs. As a proof of concept, we showed that these markers can be used to separate sainfoin individuals based on their origin.

Relationship between modified Roger’s Distance to Euclidian Distance and to Nei’s Distance. (PDF 319 kb)

Background Sainfoin ( Onobrychis viciifolia ) is a highly nutritious tannin-containing forage legume. In the diet of ruminants sainfoin can have anti-parasitic effects and reduce methane emissions under in vitro conditions. Many of these benefits have been attributed to condensed tannins or proanthocyanidins in sainfoin. A combination of increased use of industrially produced nitrogen fertilizer, issues with establishment and productivity in the first year and more reliable alternatives, such as red clover led to a decline in the use of sainfoin since the middle of the last century. In recent years there has been a resurgence of interest in sainfoin due to its potential beneficial nutraceutical and environmental attributes. However, genomic resources are scarce, thus hampering progress in genetic analysis and improvement. To address this we have used next generation RNA sequencing technology to obtain the first transcriptome of sainfoin. We used the library to identify gene-based simple sequence repeats (SSRs) and potential single nucleotide polymorphisms (SNPs). Results One genotype from each of five sainfoin accessions was sequenced. Paired-end (PE) sequences were generated from cDNA libraries of RNA extracted from 7 day old seedlings. A combined assembly of 92,772 transcripts was produced de novo using the Trinity programme. About 18,000 transcripts were annotated with at least one GO (gene ontology) term. A total of 63 transcripts were annotated as involved in the tannin biosynthesis pathway. We identified 3786 potential SSRs. SNPs were identified by mapping the reads of the individual assemblies against the combined assembly. After stringent filtering a total of 77,000 putative SNPs were identified. A phylogenetic analysis of single copy number genes showed that sainfoin was most closely related to red clover and Medicago truncatula , while Lotus japonicus , bean and soybean are more distant relatives. Conclusions This work describes the first transcriptome assembly in sainfoin. The 92 K transcripts provide a rich source of SNP and SSR polymorphisms for future use in genetic studies of this crop. Annotation of genes involved in the condensed tannin biosynthesis pathway has provided the basis for further studies of the genetic control of this important trait in sainfoin.

Emmer wheat, Triticum dicoccon Schrank was one of the founder crops of Neolithic agriculture. Though its cultivation was largely replaced by hexaploid wheats 2000 years ago, pockets of small scale cultivation can still be found. One such area is the Asturias region of Northern Spain, where emmer wheat remains a traditional crop for high value specialist culinary uses, and farmers grow locally adapted landraces. In order to study the diversity of these landraces, we sampled emmer wheat from different regions of Asturias, and genotyped multiple plants from each village using nuclear and chloroplast microsatellites. A high level of variation was observed with markers from both genomes, including a novel chloroplast haplotype. A strong geographic structure was observed in the Asturian emmer wheats in both the chloroplast markers and the nuclear microsatellite data.

Sainfoin (Onobrychis viciifolia Scop.) is a perennial forage legume that possesses beneficial properties in the context of sustainable agriculture. A germplasm collection of O. viciifolia was assembled from many geographic regions, as well as a collection of Onobrychis species that might be crossed with O. viciifolia to improve its biological and agronomic properties. The objective of the present study was to obtain cytological information for a broad range of O. viciifolia varieties and related species, in order to confirm and extend previous findings. The study set was 143 accessions of O. viciifolia as well as 34 accessions of 17 other species from the Onobrychis genus. Three of the O. viciifolia accessions were diploid (2n = 2x = 14) and the remainder tetraploid (2n = 4x = 28). Chromosomes were <5 μm in length. Among the other Onobrychis species, roughly half were found to be diploid and the other half tetraploid, with two species, O. arenaria (Kit.) DC. and O. alba subsp. laconica (Orph. ex Boiss.) Hayek, including a mixture of diploid and tetraploid accessions. The basic number of chromosomes was seven, except for O. aequidentata (Sm.) d’Urv. and O. crista-galli Lam., where it was eight. The 2C value for O. viciifolia was estimated at 2.5 pg. These cytological data will be valuable as part of a pre-breeding programme aimed at the eventual production of improved O. viciifolia varieties for use in sustainable agriculture.

Results about a phylogenetic analysis of the genus Onobrychis Mill., tribe Hedysareae DC. are presented. The systematic knowledge of tribe Hedysareae is still incomplete, with difficult circumscription of genera and species. Analyses were undertaken using both nuclear (ITS) and chloroplast (matK) markers for a set of 78 accessions covering 41 Onobrychis species, besides previously sequenced Hedysareae accessions. The phylogenetic methods used were maximum parsimony, maximum likelihood, and Bayesian analyses to produce phylogenetic trees and robustness indices. The genus Onobrychis was resolved as paraphyletic, with species of the genera Eversmannia Bunge and Hedysarum L. nested within it. The position of the section Membranacea of genus Hedysarum was as a sister group to Onobrychis and Eversmannia, separated from other accessions of Hedysarum. Variation in the 2 markers was sufficient to resolve infrageneric groups in Onobrychis and Hedysarum, but we were unable to completely resolve certain species in Onobrychis, particularly those within the sect. Onobrychis. The cause of this difficult species delimitation may be related to recent speciation, hybridization, and introgression events, particularly between cultivated species and their wild relatives, and the presence of cryptospecies as suggested by intraspecific polyploid series.

Results of analyses of the photoperiod response gene (PPD-H1) and simple sequence repeats (SSRs) in modern landraces of cultivated barley were used as evidence for the mechanism of agricultural spread in Neolithic Europe. In particular, we explored the usefulness of considering adaptive genes as indicators of past selective pressures acting on crops, during their spread through Europe. In some areas, such as the Alpine region, Britain and Scandinavia, we have evidence to suggest that the adaptation of crops to certain climatic conditions may have contributed to the timing of agricultural spread. At the northern fringes of Europe, and in higher altitude locations in central Europe, the introduction of more suitably adapted cereals may have facilitated successful agriculture to trigger agricultural expansion. This research opens up the possibility of investigating other genetic adaptations to climate, which would permit a fuller evaluation of the relative contributions of climate/crop and forager/farmer interactions in the process of agricultural spread.