The functional role of methylated short tandem repeats in early mouse development

Short tandem repeats, or microsatellites are ubiquitous throughout all genomes that have been explored. In common with other sequences, the DNA in microsatellites has DNA marks in the form of chromatin methylation. Regulation of DNA methylation and changes in their pattern is critical for the establishment of unique cell states throughout development in mammals. DNA methylation is extensively reprogrammed during the early phases of mammalian development to establish unique developmental patterning. Whether microsatellites are also reprogrammed with developmental patterns is unknown. In this thesis, we assessed the characteristics of di- and trinucleotide microsatellites in the NCBIM37 Mus musculus assembly and observed a marked difference in quantity and length of microsatellites of differing motif, not explained by any known mechanism. Secondly we assessed the quantities of di-, tri- and tetranucleotide microsatellites in experimentally determined methylomes of Mus musculus at various stages in development. Our results indicate that at least one tetranucleotide microsatellite motif and more tentatively a second trinucleotide microsatellite follow a pattern of methylation consistent with reprogramming. Finally we show that the genes containing these specific microsatellites in the NCBIM37 genome have strong links to known developmental processes.Biotechnology and Biological (BBSRC)Applied Bioinformatic

Light spectra blocking films reduce numbers of western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae) in strawberry, Fragaria x ananassa

Frankliniella occidentalis is a pest of horticultural crops, including commercial strawberry (Fragaria x ananassa). Control is challenging because certain populations are resistant to insecticides and, in strawberry, now relies on the application of biocontrols. However, this approach is not always successful if F. occidentalis populations overwhelm biocontrols. We investigated whether targeted spectral modifications to cladding materials could reduce numbers of F. occidentalis, in strawberry flowers. Five UV-attenuating plastic-film materials were tested in three, 6-week, semi-field tunnel experiments containing strawberry plants. F. occidentalis were introduced into tunnels from a laboratory culture and subsequent numbers that developed in strawberry flowers were recorded. Limiting UV-A radiation to the crop significantly reduced the numbers of adult and larval F. occidentalis in strawberry flowers. The numbers of adult (and larvae) in flowers were reduced by 42 (47)%, 54 (41)%, 70 (73)%, and 82 (73)% in UV350, UV370, UV400, and UV430-attenuating films, respectively, compared with the UVopen (control) film. However, no damage to strawberry fruits was observed regardless of the film treatment. Incorporating UV-attenuating films as tunnel cladding can suppress F. occidentalis numbers in strawberry. Reducing populations of F. occidentalis in crops is likely to enable the more successful use of other non-chemical control strategies

Investigating the inoculum dynamics of Cladosporium on the surface of raspberry fruits and in the air

Raspberry production is under threat from the emerging fungal pathogenic genus Cladosporium. We used amplicon-sequencing, coupled with qPCR, to investigate how fruit age, fruit location within a polytunnel, polytunnel location and sampling date affected the fruit epiphytic microbiome. Fruit age was the most important factor impacting the fungal microbiome, followed by sampling date and polytunnel location. In contrast, polytunnel location and fruit age were important factors impacting the bacterial microbiome composition, followed by the sampling date. The within-tunnel location had a small significant effect on the fungal microbiome and no effect on the bacterial microbiome. As fruit ripened, fungal diversity increased and the bacterial diversity decreased. Cladosporium was the most abundant fungus of the fruit epiphytic microbiome, accounting for nearly 44% of all fungal sequences. Rotorod air samplers were used to study how the concentration of airborne Cladosporium inoculum (quantified by qPCR) varied between location (inside and outside the polytunnel) and time (daytime vs. nighttime). Quantified Cladosporium DNA was significantly higher during the day than the night and inside the polytunnel than the outside. This study demonstrated the dynamic nature of epiphytic raspberry fruit microbiomes and airborne Cladosporium inoculum within polytunnels, which will impact disease risks on raspberry fruit.Biotechnology and Biological Sciences Research Council, Grant/Award Number: BB/T509073/

Viral interactions and pathogenesis during multiple viral infections in Agaricus bisporus

Viral interactions during multiple viral infections were examined in Agaricus bisporus cultures harboring 9 viruses (comprising 18 distinct viral RNAs) by statistically analyzing their relative abundance in fruitbodies. Four clusters of viral RNA were identified that suggested synergism and coreplication. Pairwise correlations revealed negative and positive correlations between clusters, indicating further synergisms and an antagonism involving a group containing a putative hypovirus and four nonhost ORFan RNAs (RNAs with no similarity to known sequences) possibly acting as defective interfering RNAs. The disease phenotype was observed in 10 to 15% of the fruitbodies apparently randomly located among asymptomatic fruitbodies. The degree of symptom expression consistently correlated with the levels of the multipartite virus AbV16. Diseased fruitbodies contained very high levels of AbV16 and AbV6 RNA2; these levels were orders of magnitude higher than those in asymptomatic tissues and were shown statistically to be discretely higher populations of abundance, indicating an exponential shift in the replicative capacity of the virus. High levels of AbV16 replication were specific to the fruitbody and not found in the underlying mycelium. There appeared to be a stochastic element occurring in these viral interactions, as observed in the distribution of diseased symptoms across a culture, differences in variance between experiments, and a number of additional viruses undergoing the step-jump in levels between experiments. Possible mechanisms for these multiple and simultaneous viral interactions in single culture are discussed in relation to known virus-host regulatory mechanisms for viral replication and whether additional factors could be considered to account for the 1,000-fold increase in AbV16 and AbV6 RNA2 levels. IMPORTANCE How viruses interact in a multiple-virus infection was examined by quantifying the levels of 18 viral RNAs in fruiting cultures of the agriculturally cultivated fungus Agaricus bisporus and statistically analyzing and modeling their abundance. Synergistic, antagonistic, and neutral interactions occurred simultaneously in cultures. The viral RNAs were grouped into four clusters, each displaying similar relative abundance, and between clusters, further interactions were found with positive, negative, or no correlations. Mushroom fruitbodies showing disease symptoms were distributed apparently randomly across the culture. These symptoms were associated with the presence of viral RNAs from two different clusters at very high levels, 1,000-fold higher than asymptomatic fruitbodies. The role of viral interaction together with stochastic factors and the regulation of host antiviral defenses in pathogenesis are discussed

CropQuant-Air: an AI-powered system to enable phenotypic analysis of yield- and performance-related traits using wheat canopy imagery collected by low-cost drones

As one of the most consumed stable foods around the world, wheat plays a crucial role in ensuring global food security. The ability to quantify key yield components under complex field conditions can help breeders and researchers assess wheat’s yield performance effectively. Nevertheless, it is still challenging to conduct large-scale phenotyping to analyse canopy-level wheat spikes and relevant performance traits, in the field and in an automated manner. Here, we present CropQuant-Air, an AI-powered software system that combines state-of-the-art deep learning (DL) models and image processing algorithms to enable the detection of wheat spikes and phenotypic analysis using wheat canopy images acquired by low-cost drones. The system includes the YOLACT-Plot model for plot segmentation, an optimised YOLOv7 model for quantifying the spike number per m2 (SNpM2) trait, and performance-related trait analysis using spectral and texture features at the canopy level. Besides using our labelled dataset for model training, we also employed the Global Wheat Head Detection dataset to incorporate varietal features into the DL models, facilitating us to perform reliable yield-based analysis from hundreds of varieties selected from main wheat production regions in China. Finally, we employed the SNpM2 and performance traits to develop a yield classification model using the Extreme Gradient Boosting (XGBoost) ensemble and obtained significant positive correlations between the computational analysis results and manual scoring, indicating the reliability of CropQuant-Air. To ensure that our work could reach wider researchers, we created a graphical user interface for CropQuant-Air, so that non-expert users could readily use our work. We believe that our work represents valuable advances in yield-based field phenotyping and phenotypic analysis, providing useful and reliable toolkits to enable breeders, researchers, growers, and farmers to assess crop-yield performance in a cost-effective approach

Amplicon sequencing identified a putative pathogen, Macrophomina phaseolina, causing wilt in African eggplant (Solanum aethiopicum) grown in Tanzania and Uganda

African eggplant (Solanum aethiopicum L.) is one of the most common traditional vegetables in Tanzania and Uganda, but its productivity is severely affected by wilt diseases caused by a number of pathogens. Plant stem and root samples were collected in several fields from many neighboring diseased and healthy plants of the Gilo group in Tanzania and from the Shum group in Uganda to identify putative pathogens causing wilt on African eggplants. Through amplicon sequencing of sampled diseased and healthy tissues, we identified putative causal pathogens for the wilt symptoms. Wilting of S. aethiopicum in Uganda is most likely caused by the bacterial pathogen Ralstonia solanacearum whereas, in Tanzania, wilt is most likely caused by the fungal pathogen Macrophomina phaseolina, infecting roots. Infection of stems by Fusarium solani may also contribute to the wilt symptoms in Tanzania. Further artificial inoculation under controlled conditions confirmed that M. phaseolina can cause typical wilting symptoms on S. aethiopcium genotypes. The discovery of different putative causal agents of wilt in the crop demonstrates the need for site specific etiological analysis of wilt before developing and implementing effective control methods. Further research is needed to confirm the results and develop appropriate management measures against specific wilt pathogens

Learning Progression: Implications for Curriculum and Assessment

This short brief is targeted to policy makers in governments and school districts who may be thinking about learning progressions and the implications for curriculum, teaching, and assessment

Oral abstracts 1: SpondyloarthropathiesO1. Detecting axial spondyloarthritis amongst primary care back pain referrals

Background: Inflammatory back pain (IBP) is an early feature of ankylosing spondylitis (AS) and its detection offers the prospect of early diagnosis of AS. However, since back pain is very common but only a very small minority of back pain sufferers have ASpA or AS, screening of back pain sufferers for AS is problematic. In early disease radiographs are often normal so that fulfilment of diagnostic criteria for AS is impossible though a diagnosis of axial SpA can be made if MRI evidence of sacroiliitis is present. This pilot study was designed to indicate whether a cost-effective pick up rate for ASpA/early AS could be achieved by identifying adults with IBP stratified on the basis of age. Methods: Patients aged between 18 and 45 years who were referred to a hospital physiotherapy service with back pain of more than 3 months duration were assessed for IBP. All were asked to complete a questionnaire based on the Berlin IBP criteria. Those who fulfilled IBP criteria were also asked to complete a second short questionnaire enquiring about SpA comorbidities, to have a blood test for HLA-B27 and CRP level and to undergo an MRI scan of the sacroiliac joints. This was a limited scan, using STIR, diffusion-weighted, T1 and T2 sequences of the sacroiliac joints to minimize time in the scanner and cost. The study was funded by a research grant from Abbott Laboratories Ltd. Results: 50 sequential patients agreed to participate in the study and completed the IBP questionnaire. Of these 27 (54%) fulfilled criteria for IBP. Of these, 2 patients reported a history of an SpA comorbidity - 1 psoriasis; 1 ulcerative colitis - and 3 reported a family history of an SpA comorbidity - 2 psoriasis; 1 Crohn's disease. 4 were HLA-B27 positive, though results were not available for 7. Two patients had marginally raised CRP levels (6, 10 -NR ≤ 5). 19 agreed to undergo MRI scanning of the sacroiliac joints and lumbar spine; 4 scans were abnormal, showing evidence of bilateral sacroiliitis on STIR sequences. In all cases the changes met ASAS criteria but were limited. Of these 4 patients 3 were HLA-B27 positive but none gave a personal or family history of an SpA-associated comorbidity and all had normal CRP levels. Conclusions: This was a pilot study yielding only limited conclusions. However, it is clear that: Screening of patients referred for physiotherapy for IBP is straightforward, inexpensive and quick. It appears that IBP is more prevalent in young adults than overall population data suggest so that targeting this population may be efficient. IBP questionnaires could be administered routinely during a physiotherapy assessment. HLA-B27 testing in this group of patients with IBP is a suitable screening tool. The sacroiliac joint changes identified were mild and their prognostic significance is not yet clear so that the value of early screening needs further evaluation. Disclosure statement: C.H. received research funding for this study from Abbott. A.K. received research funding for this study, and speaker and consultancy fees, from Abbott. All other authors have declared no conflicts of interes