Progress and opportunities through use of genomics in animal production.

The rearing of farmed animals is a vital component of global food production systems, but its impact on the environment, human health, animal welfare, and biodiversity is being increasingly challenged. Developments in genetic and genomic technologies have had a key role in improving the productivity of farmed animals for decades. Advances in genome sequencing, annotation, and editing offer a means not only to continue that trend, but also, when combined with advanced data collection, analytics, cloud computing, appropriate infrastructure, and regulation, to take precision livestock farming (PLF) and conservation to an advanced level. Such an approach could generate substantial additional benefits in terms of reducing use of resources, health treatments, and environmental impact, while also improving animal health and welfare.

Modulation of physio-biochemical and photosynthesis parameters by overexpressing SbPIP2 gene improved abiotic stress tolerance of transgenic tobacco.

The present study aims to explore the potential of a plasma-membrane localized PIP2-type aquaporin protein sourced from the halophyte Salicornia brachiata to alleviate salinity and water deficit stress tolerance in a model plant through transgenic intervention. Transgenic plants overexpressing SbPIP2 gene showed improved physio-biochemical parameters like increased osmolytes (proline, total sugar, and amino acids), antioxidants (polyphenols), pigments and membrane stability under salinity and drought stresses compared to control plants [wild type (WT) and vector control (VC) plants]. Multivariate statistical analysis showed that, under water and salinity stresses, osmolytes, antioxidants and pigments were correlated with SbPIP2-overexpressing (SbPIP2-OE) plants treated with salinity and water deficit stress, suggesting their involvement in stress tolerance. As aquaporins are also involved in CO2 transport, SbPIP2-OE plants showed enhanced photosynthesis performance than wild type upon salinity and drought stresses. Photosynthetic gas exchange (net CO2 assimilation rate, PSII efficiency, ETR, and non-photochemical quenching) were significantly higher in SbPIP2-OE plants compared to control plants (wild type and vector control plants) under both unstressed and stressed conditions. The higher quantum yield for reduction of end electron acceptors at the PSI acceptor side [Φ( R0 )] in SbPIP2-OE plants compared to control plants under abiotic stresses indicates a continued PSI functioning, leading to retained electron transport rate, higher carbon assimilation, and less ROS-mediated injuries. In conclusion, the SbPIP2 gene functionally validated in the present study could be a potential candidate for engineering abiotic stress resilience in important crops.

Assessing student perception of the integration of portable wireless ultrasound imaging in undergraduate anatomy education.

Anatomy is the foundation of many physiology and healthcare-related degrees. With limited access to cadavers in many universities, it is essential to investigate techniques that could be utilized to support and enhance the teaching of anatomy. Ultrasound is used clinically to aid the diagnosis of a wide range of conditions by visualizing the anatomy of the patient. While research has investigated the advantages of ultrasound in medical education, the potential benefits of ultrasound in undergraduate bioscience degrees remain to be investigated. The aim of this study was to identify if a portable ultrasound probe that wirelessly attaches to a smartphone or tablet was perceived by students as beneficial for their understanding and learning of anatomy, and to identify if there were any barriers for students partaking in ultrasound sessions. Following five ultrasound-teaching sessions, 107 undergraduate students completed a 5-point likert questionnaire on their perception of the integration of portable ultrasound machines in anatomy education. The data indicated that 93% of students perceived that the ultrasound teaching sessions improved their anatomical understanding, 94% perceived that ultrasound increased their ability to understand the clinical relevance of learning anatomy, 97% enjoyed the sessions, and 95% of students believed that ultrasound should be integrated into anatomy teaching. In this study, we also found several barriers for students taking part in ultrasound sessions, including religious beliefs, and lacking adequate background knowledge. In conclusion, these findings demonstrate, for the first time, that students perceive portable ultrasound to enhance their anatomy studies, demonstrating the potential benefit the integration of ultrasound into the anatomy curriculum may serve within undergraduate bioscience courses.

Co-Loading of Black Phosphorus Nanoflakes and Doxorubicin in Lysolipid Temperature-Sensitive Liposomes for Combination Therapy in Prostate Cancer.

Black phosphorus (BP) is one of the most promising nanomaterials for cancer therapy. This 2D material is biocompatible and has strong photocatalytic activity, making it a powerful photosensitiser for combined NIR photothermal and photodynamic therapies. However, the fast degradation of BP in oxic conditions (including biological environments) still limits its use in cancer therapy. This work proposes a facile strategy to produce stable and highly concentrated BP suspensions using lysolipid temperature-sensitive liposomes (LTSLs). This approach also allows for co-encapsulating BP nanoflakes and doxorubicin, a potent chemotherapeutic drug. Finally, we demonstrate that our BP/doxorubicin formulation shows per se high antiproliferative action against an in vitro prostate cancer model and that the anticancer activity can be enhanced through NIR irradiance.

A novel technology of solarization and phytoremediation enhanced with biosurfactant for the sustainable treatment of PAH-contaminated soil.

Hydrocarbon-contaminated land has been a significant issue throughout Nigeria's Niger Delta since the discovery of crude oil in 1956. This paper proposes a novel and sustainable technique involving soil solarization, phytoremediation and biosurfactant to treat polycyclic aromatic hydrocarbon (PAH) contamination. The treatment effect on PAH reduction, plant growth, rhizosphere microorganisms and their enzymatic activities was evaluated. Twenty-eight days of solarization was carried out before the introduction of Chromolaena odorata seedlings for an 84-day phytoremediation period using a 4 × 4 (vegetated) and 2 × 4 (non-vegetated) cell microcosms to simulate the Niger Delta's subtropical conditions. Soil solarization resulted in significant PAH reduction (p ≤ 0.01) of phenanthrene, fluoranthene and benzo(a)pyrene with means reduction of 60.0%, 38.7% and 36.1% compared to their non-solarized counterparts with 18.0%, 18.0% and 18.8% at 95% CI (32.7, 51.3), (15.4, 26.1) and (8.0, 26.6), respectively. In post-solarization, all solarized and vegetated treatment groups significantly reduced (p ≤ 0.01) PAHs compared to their respective counterparts, while biosurfactant contribution in this combination was negligible (p ≥ 0.05). The growth parameters of C. odorata, total soil/rhizosphere heterotrophic microorganisms and their enzymatic activities of dehydrogenase and urease increased in all solarized treatments indicating essential biochemical processes. The novel and successful integration of soil solarization and phytoremediation using indigenous C. odorata as a combined technique to treat even the most recalcitrant form of hydrocarbons (PAHs) brings up new opportunities for a sustainable method of cleaning up contaminated land in Nigeria's oil-rich Niger Delta.

Nutritional and genetic variation in a core set of Ethiopian Tef (Eragrostis tef) varieties.

BACKGROUND: Tef (Eragrostis tef) is a tropical cereal domesticated and grown in the Ethiopian highlands, where it has been a staple food of Ethiopians for many centuries. Food insecurity and nutrient deficiencies are major problems in the country, so breeding for enhanced nutritional traits, such as Zn content, could help to alleviate problems with malnutrition. RESULTS: To understand the breeding potential of nutritional traits in tef a core set of 24 varieties were sequenced and their mineral content, levels of phytate and protein, as well as a number of nutritionally valuable phenolic compounds measured in grain. Significant variation in all these traits was found between varieties. Genome wide sequencing of the 24 tef varieties revealed 3,193,582 unique SNPs and 897,272 unique INDELs relative to the tef reference var. Dabbi. Sequence analysis of two key transporter families involved in the uptake and transport of Zn by the plant led to the identification of 32 Zinc Iron Permease (ZIP) transporters and 14 Heavy Metal Associated (HMA) transporters in tef. Further analysis identified numerous variants, of which 14.6% of EtZIP and 12.4% of EtHMA variants were non-synonymous changes. Analysis of a key enzyme in flavanol synthesis, flavonoid 3'-hydroxylase (F3'H), identified a T-G variant in the tef homologue Et_s3159-0.29-1.mrna1 that was associated with the differences observed in kaempferol glycoside and quercetin glycoside levels. CONCLUSION: Wide genetic and phenotypic variation was found in 24 Ethiopian tef varieties which would allow for breeding gains in many nutritional traits of importance to human health.

Pharmacologically induced weight loss is associated with distinct gut microbiome changes in obese rats.

BACKGROUND: Obesity, metabolic disease and some psychiatric conditions are associated with changes to relative abundance of bacterial species and specific genes in the faecal microbiome. Little is known about the impact of pharmacologically induced weight loss on distinct microbiome species and their respective gene programs in obese individuals. METHODOLOGY: Using shotgun metagenomics, the composition of the microbiome was obtained for two cohorts of obese female Wistar rats (n = 10-12, total of 82) maintained on a high fat diet before and after a 42-day treatment with a panel of four investigatory or approved anti-obesity drugs (tacrolimus/FK506, bupropion, naltrexone and sibutramine), alone or in combination. RESULTS: Only sibutramine treatment induced consistent weight loss and improved glycaemic control in the obese rats. Weight loss was associated with reduced food intake and changes to the faecal microbiome in multiple microbial taxa, genes, and pathways. These include increased ß-diversity, increased relative abundance of multiple Bacteroides species, increased Bacteroides/Firmicutes ratio and changes to abundance of genes and species associated with obesity-induced inflammation, particularly those encoding components of the flagellum and its assembly. CONCLUSIONS: Sibutramine-induced weight loss in obese rats is associated with improved metabolic health, and changes to the faecal microbiome consistent with a reduction in obesity-induced bacterially-driven inflammation.

Impact of the COVID-19 pandemic on acute otolaryngology inpatient activity at a tertiary referral centre: A retrospective analysis.

OBJECTIVE: To determine the impact of the COVID-19 pandemic on acute admissions and inpatient activity at a tertiary referral centre. DESIGN: Retrospective review of coding-based inpatient electronic records. SETTING: An otolaryngology and head and neck surgery department at a major UK trauma and tertiary referral centre. PARTICIPANTS: Otolaryngology patients admitted as an emergency over a period of 10 months pre-COVID19 (01/04/2019-23/01/2020) and 10 months post-COVID19 (01/04/2020-23/01/2021). MAIN OUTCOME MEASURES: Baseline characteristics, admission rates, length of stay (LoS), overall mortality and 30-day mortality. RESULTS: A total of 1620 records were reviewed; (1066 pre-COVID19, 554 post-COVID19). Admissions across all age groups were reduced, with an increase in mean age from 39.88 to 47.4 years (p = .018). LoS remained unchanged (3.85 vs 3.82 days, p = .160). Infection remained the most common presentation, followed secondly by epistaxis which entailed an increased LoS compared to the pre-COVID19 cohort. GP referrals reduced from 18.3% to 4.2% (n = 195 vs n = 23, p < .001) and ED referrals proportionally increased from 71.9% to 85.9% (n = 766 vs n = 476, p < .001). Critical care admissions were higher in the post-COVID19 cohort (OR 1.78 (1.07-2.98) [95% CI], p = .027). There was no significant difference in overall mortality between groups (n = 60, 5.6% vs. n = 33, 6.0%; p = .844). Thirty-day mortality increased from 0.9% (n = 12) pre-COVID19 to 2.3% (n = 13) post-COVID19 (p = .039). CONCLUSIONS: This study demonstrates significant changes and a reduction in acute otolaryngology presentations. Our findings may suggest that sicker, frailer patients were admitted during the pandemic. This study reports the observational effect that the pandemic has had on acute otolaryngology admissions, which may be relevant in addressing unmet care needs in the post-pandemic period.

Future-proofing regulation for rapidly changing biotechnologies.

Our understanding of DNA structure and how it interacts with the environment to give form and function at the organism level is growing at an unprecedented pace which shows no sign of slowing. These developments have already led to many new products and will continue to underpin as yet unpredicted future developments in biotechnology. However, this potential will not be realised unless the mechanisms for risk assessment, regulatory approval, product claims and labelling etc. are fit for purpose, have the confidence of all stakeholders and are sufficiently agile to support this rapidly changing field. The sectors that are making particular advances in biotechnological processes include agriculture, pharmaceuticals, food, chemical and human diagnostics and therapeutics. In many of these areas the research, investment and innovation pipeline is operating well as evidenced by the many marketed products. However, developments in plant breeding methods have posed particular challenges for regulators which in turn is stifling R&D and innovation, particularly in the EU. In rapidly moving areas of research and development, it is imperative that regulatory frameworks are future-proofed by design.

Prosthetic mesh placement for the prevention of parastomal herniation.

BACKGROUND: Parastomal herniation is a common problem following formation of a stoma after both elective and emergency abdominal surgery. Symptomatic hernias give rise to a significant amount of patient morbidity, and in some cases mortality, and therefore may necessitate surgical treatment to repair the hernial defect and/or re-site the stoma. In an effort to reduce this complication, recent research has focused on the application of a synthetic or biological mesh, inserted during stoma formation to help strengthen the abdominal wall. OBJECTIVES: The primary objective was to evaluate whether mesh reinforcement during stoma formation reduces the incidence of parastomal herniation. Secondary objectives included the safety or potential harms or both of mesh placement in terms of stoma-related infections, mesh-related infections, patient-reported symptoms/postoperative quality of life, and re-hospitalisation/ambulatory visits. SEARCH METHODS: We searched the Cochrane Central Register of Controlled Trials (CENTRAL; the Cochrane Library 2018, Issue 1), Ovid MEDLINE (1970 to 11 January 2018), Ovid Embase (1974 to 11 January 2018), and Science Citation Index Expanded (1970 to 11 January 2018). To identify ongoing studies, we also searched the metaRegister of Controlled Trials (mRCT) on 11 January 2018. SELECTION CRITERIA: We considered for inclusion all randomised controlled trials (RCTs) of prosthetic mesh (including biological/composite mesh) placement versus a control group (no mesh) for the prevention of parastomal hernia. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed the studies identified by the literature search for potential eligibility. We obtained the full articles for all studies that potentially met the inclusion criteria and included all those that met the criteria. Any differences in opinion between review authors were resolved by consensus. We pooled study data into a meta-analysis. We assessed heterogeneity by calculation of I2 and expressed results for each variable as a risk ratio (RR) with corresponding 95% confidence intervals (CI). We expressed continous outcomes as mean difference (MD) with corresponding 95% CIs. MAIN RESULTS: We included 10 RCTs involving a total of 844 participants. The primary outcome was overall incidence of parastomal herniation. Secondary outcomes were rate of reoperation at 12 months, operative time, postoperative length of hospital stay, stoma-related infections, mesh-related infections, quality of life, and rehospitalisation rate. We judged the risk of bias across all domains to be low in six trials. We judged four trials to have an overall high risk of bias.The overall incidence of parastomal hernia was less in participants receiving a prophylactic mesh compared to those who had a standard ostomy formation (RR 0.53, 95% CI 0.43 to 0.66; 10 studies, 771 participants; I2 = 69%; low-quality evidence). In absolute numbers, the incidence of parastomal hernia was 22 per 100 participants (18 to 27) receiving prophylactic mesh compared to 41 per 100 participants having a standard ostomy formation.There were no differences in the need for reoperation (RR 0.90, 95% CI 0.50 to 1.64; 9 studies, 757 participants; I2 = 0%; low-quality evidence); operative time (MD -6.50 (min), 95% CI -18.24 to 5.24; 6 studies, 671 participants; low-quality evidence); postoperative length of hospital stay (MD -0.95 (days), 95% CI -2.03 to 0.70; 4 studies, 500 participants; moderate-quality evidence); or stoma-related infections (RR 0.89, 95% CI 0.32 to 2.50; 6 studies, 472 participants; I2 = 0%; low-quality evidence) between the two groups.We were unable to analyse mesh-related infections, quality of life, and rehospitalisation rate due to sparse data or because the outcome was not reported in the included studies. AUTHORS' CONCLUSIONS: This Cochrane Review included 10 RCTs with a total of 844 participants. The review demonstrated a reduction in the incidence of parastomal hernia in people who had a prophylactic synthetic mesh placed at the time of the index operation compared to a standard ostomy formation. However, our confidence in this estimate is low due to the presence of a large degree of clinical heterogeneity, as well as high variability in follow-up duration and technique of parastomal herniation detection. We found the rate of stoma-related infection to be similar in both the intervention and control groups.

Oxidized LDL activates blood platelets through CD36/NOX2-mediated inhibition of the cGMP/protein kinase G signaling cascade.

Oxidized low-density lipoprotein (oxLDL) promotes unregulated platelet activation in dyslipidemic disorders. Although oxLDL stimulates activatory signaling, it is unclear how these events drive accelerated thrombosis. Here, we describe a mechanism for oxLDL-mediated platelet hyperactivity that requires generation of reactive oxygen species (ROS). Under arterial flow, oxLDL triggered sustained generation of platelet intracellular ROS, which was blocked by CD36 inhibitors, mimicked by CD36-specific oxidized phospholipids, and ablated in CD36(-/-) murine platelets. oxLDL-induced ROS generation was blocked by the reduced NAD phosphate oxidase 2 (NOX2) inhibitor, gp91ds-tat, and absent in NOX2(-/-) mice. The synthesis of ROS by oxLDL/CD36 required Src-family kinases and protein kinase C (PKC)-dependent phosphorylation and activation of NOX2. In functional assays, oxLDL abolished guanosine 3',5'-cyclic monophosphate (cGMP)-mediated signaling and inhibited platelet aggregation and arrest under flow. This was prevented by either pharmacologic inhibition of NOX2 in human platelets or genetic ablation of NOX2 in murine platelets. Platelets from hyperlipidemic mice were also found to have a diminished sensitivity to cGMP when tested ex vivo, a phenotype that was corrected by infusion of gp91ds-tat into the mice. This study demonstrates that oxLDL and hyperlipidemia stimulate the generation of NOX2-derived ROS through a CD36-PKC pathway and may promote platelet hyperactivity through modulation of cGMP signaling.

Genetic and Epigenetic Intra-tumour Heterogeneity in Colorectal Cancer.

INTRODUCTION: Colorectal cancer (CRC) is a highly heterogeneous disease, with pathologically similar cancers having completely different responses to treatment and patient survival. Intra-tumour heterogeneity (defined as distinct morphological and phenotypic differences) has recently been demonstrated to be an important factor in the development and behaviour of cancer cells and can be used to determine response to anticancer therapy. METHOD: Patients with resected CRC had DNA extracted from eight defined tumour areas which were analysed for two genetic mutations (BRAF and KRAS) and one epigenetic trait (CpG island methylator phenotype/CIMP). Normal adjacent tissue was studied as control. RESULTS: Twelve patients with CRC were included. Intra-tumoural heterogeneity for KRAS mutation was seen in 2 patients (17%). There was no statistical evidence of CIMP status heterogeneity (p = 0.85), but 6 of the 12 patients (50%) demonstrated at least one heterogeneous area within the tumour. DISCUSSION: Intra-tumoural heterogeneity for both genetic and epigenetic factors in CRC is more prevalent than previously thought in Stage II and Stage III CRC. This study provides new insight into epigenetic heterogeneity of CRC and supports the development of a more targeted biopsy strategy to support expansion of personalised treatment.

Acute liver effects, disposition and metabolic fate of [14C]-fenclozic acid following oral administration to normal and bile-cannulated male C57BL/6J mice.

The distribution, metabolism, excretion and hepatic effects of the human hepatotoxin fenclozic acid were investigated following single oral doses of 10 mg/kg to normal and bile duct-cannulated male C57BL/6J mice. Whole body autoradiography showed distribution into all tissues except the brain, with radioactivity still detectable in blood, kidney and liver at 72 h post-dose. Mice dosed with [14C]-fenclozic acid showed acute centrilobular hepatocellular necrosis, but no other regions of the liver were affected. The majority of the [14C]-fenclozic acid-related material recovered was found in the urine/aqueous cage wash, (49%) whilst a smaller portion (13%) was eliminated via the faeces. Metabolic profiles for urine, bile and faecal extracts, obtained using liquid chromatography and a combination of mass spectrometric and radioactivity detection, revealed extensive metabolism of fenclozic acid in mice that involved biotransformations via both oxidation and conjugation. These profiling studies also revealed the presence of glutathione-derived metabolites providing evidence for the production of reactive species by mice administered fenclozic acid. Covalent binding to proteins from liver, kidney and plasma was also demonstrated, although this binding was relatively low (less than 50 pmol eq./mg protein).

Hepatic effects of repeated oral administration of diclofenac to hepatic cytochrome P450 reductase null (HRN™) and wild-type mice.

Hepatic NADPH-cytochrome P450 oxidoreductase null (HRN™) mice exhibit normal hepatic and extrahepatic biotransformation enzyme activities when compared to wild-type (WT) mice, but express no functional hepatic cytochrome P450 activities. When incubated in vitro with [(14)C]-diclofenac, liver microsomes from WT mice exhibited extensive biotransformation to oxidative and glucuronide metabolites and covalent binding to proteins was also observed. In contrast, whereas glucuronide conjugates and a quinone-imine metabolite were formed when [(14)C]-diclofenac was incubated with HRN™ mouse liver, only small quantities of P450-derived oxidative metabolites were produced in these samples and covalent binding to proteins was not observed. Livers from vehicle-treated HRN™ mice exhibited enhanced lipid accumulation, bile duct proliferation, hepatocellular degeneration and necrosis and inflammatory cell infiltration, which were not present in livers from WT mice. Elevated liver-derived alanine aminotransferase, glutamate dehydrogenase and alkaline phosphatase activities were also observed in plasma from HRN™ mice. When treated orally with diclofenac for 7 days, at 30 mg/kg/day, the severities of the abnormal liver histopathology and plasma liver enzyme findings in HRN™ mice were reduced markedly. Oral diclofenac administration did not alter the liver histopathology or elevate plasma enzyme activities of WT mice. These findings indicate that HRN™ mice are valuable for exploration of the role played by hepatic P450s in drug biotransformation, but poorly suited to investigations of drug-induced liver toxicity. Nevertheless, studies in HRN™ mice could provide novel insights into the role played by inflammation in liver injury and may aid the evaluation of new strategies for its treatment.

A critical evaluation of venous thromboembolism risk assessment models used in patients with lower limb cast immobilisation.

BACKGROUND: There is a recognised link between lower limb cast immobilisation and the development of venous thromboembolism (VTE). Our aim was to assess the diagnostic accuracy of risk assessment models (RAMs) applicable to this patient group. This has not been done before. METHODS: A literature and guideline review identified five RAMs. They were used to retrospectively risk assess a consecutive series of patients who were diagnosed with symptomatic VTE following lower limb injury treated with a cast (Group I). A case-matched cohort who did not suffer symptomatic VTE (Group II) was also retrospectively risk assessed. The RAMs' diagnostic performance indicators were calculated. RESULTS: Groups I and II consisted of 21 patients each. There was no significant difference in the mean age or total number of VTE risk factors between Groups I and II (p=.957 and p=.878 respectively). The Plymouth (2010) RAM achieved the highest accuracy (54.8%). CONCLUSIONS: Each RAM demonstrated significant limitations. Two displayed very limited clinical utility. Three recommended chemical thromboprophylaxis to all patients because they weighted lower limb immobilisation as an absolute risk factor for the development of VTE. Cast immobilisation should not be considered an absolute risk factor when risk assessing patients who all have casts. Prospective evaluation with a larger patient cohort is required.

Gluten quality of bread wheat is associated with activity of RabD GTPases.

In the developing endosperm of bread wheat (Triticum aestivum), seed storage proteins are produced on the rough endoplasmic reticulum (ER) and transported to protein bodies, specialized vacuoles for the storage of protein. The functionally important gluten proteins of wheat are transported by two distinct routes to the protein bodies where they are stored: vesicles that bud directly off the ER and transport through the Golgi. However, little is known about the processing of glutenin and gliadin proteins during these steps or the possible impact on their properties. In plants, the RabD GTPases mediate ER-to-Golgi vesicle transport. Available sequence information for Rab GTPases in Arabidopsis, rice, Brachypodium and bread wheat was compiled and compared to identify wheat RabD orthologs. Partial genetic sequences were assembled using the first draft of the Chinese Spring wheat genome. A suitable candidate gene from the RabD clade (TaRabD2a) was chosen for down-regulation by RNA interference (RNAi), and an RNAi construct was used to transform wheat plants. All four available RabD genes were shown by qRT-PCR to be down-regulated in the transgenic developing endosperm. The transgenic grain was found to produce flour with significantly altered processing properties when measured by farinograph and extensograph. SE-HPLC found that a smaller proportion of HMW-GS and large proportion of LMW-GS are incorporated into the glutenin macropolymer in the transgenic dough. Lower protein content but a similar protein profile on SDS-PAGE was seen in the transgenic grain.

Molecular characterization of two isoforms of a farnesyl pyrophosphate synthase gene in wheat and their roles in sesquiterpene synthesis and inducible defence against aphid infestation.

Aphids are important pests of wheat (Triticum aestivum) that affect crop production globally. Herbivore-induced emission of sesquiterpenes can repel pests, and farnesyl pyrophosphate synthase (FPS) is a key enzyme involved in sesquiterpene biosynthesis. However, fps orthologues in wheat and their functional roles in sesquiterpene synthesis and defence against aphid infestation are unknown. Here, two fps isoforms, Tafps1 and Tafps2, were identified in wheat. Quantitative real-time polymerase chain reaction (qRT-PCR) and in vitro catalytic activity analyses were conducted to investigate expression patterns and activity. Heterologous expression of these isoforms in Arabidopsis thaliana, virus-induced gene silencing (VIGS) in wheat and aphid behavioural assays were performed to understand the functional roles of these two isoforms. We demonstrated that Tafps1 and Tafps2 played different roles in induced responses to aphid infestation and in sesquiterpene synthesis. Heterologous expression in A. thaliana resulted in repulsion of the peach aphid (Myzus persicae). Wheat plants with these two isoforms transiently silenced were significantly attractive to grain aphid (Sitobion avenae). Our results provide new insights into induced defence against aphid herbivory in wheat, in particular, the different roles of the two Tafps isoforms in both sesquiterpene biosynthesis and defence against aphid infestation.

Implications of using genomic prediction within a high-density SNP dataset to predict DUS traits in barley.

KEY MESSAGE: Alternative methods for genomic prediction of traits and trait differences are compared and recommendations made. We make recommendations for implementing methods in the context of DUS testing. High-throughput genotyping provides an opportunity to explore the application of genotypes in predicting plant phenotypes. We use a genome-wide prediction model to estimate the contribution of all loci and sum over multiple minor effects to predict traits. A potential use is in plant variety protection to discriminate among varieties on distinctness. We investigate this use with alternate scenarios in a set of 431 winter and spring barley varieties, with trait data from UK DUS trials comprising 28 characteristics, together with SNP genotype data. Firstly, each trait is predicted from genotypes by ridge regression with discrimination among varieties using predicted traits. Secondly, squared trait differences between each pair of varieties are regressed on genetic distances between each variety by ridge regression, with discrimination among varieties using the predicted squared trait differences directly. This latter approach is analogous to the use of phenotype and marker differences introduced to human genetic linkage analysis by Haseman and Elston and to the analysis of heritability in natural populations of plants by Ritland. We compare correlations between methods, both trait by trait and summarised across all traits. Our results show wide variation among correlations for each trait. However, the aggregate distances calculated from values predicted by genotypes show higher correlations with distances calculated from measured values than any previously reported. We discuss the applicability of these results to implementation of UPOV Model 2 in DUS testing and suggest 'success criteria' that should be considered by testing authorities seeking to implement UPOV Model 2.