The influence of genetic variation and nitrogen source on nitrate accumulation and iso-osmotic regulation by lettuce

Background and aims: Characterisation of genetic variation in nitrate accumulation by lettuce will inform strategies for selecting low-nitrate varieties more capable of meeting EU legislation on harvested produce. This study uses a population of recombinant inbred lines (RILs) of lettuce to determine how genotypic differences influence N uptake, N assimilation and iso-osmotic regulation, and to identify key related traits prior to future genetic analysis. Methods: Measurements were made on plants grown to maturity in soil fertilised with ammonium nitrate, and in a complete nutrient solution containing only nitrate-N. A simple osmotic balance model was developed to estimate variations in shoot osmotic concentration between RILs. Results: There were significant genotypic variations in nitrate accumulation when plants were grown either with nitrate alone or in combination with ammonium. Ammonium-N significantly reduced nitrate in the shoot but had no effect on its relative variability, or on the ranking of genotypes. Shoot nitrate-N was correlated positively with total-N and tissue water, and negatively with assimilated-C in both experiments. Corresponding relationships with assimilated-N and shoot weight were weaker. Estimated concentrations of total osmotica in shoot sap were statistically identical in all RILs, despite variations in nitrate concentration across the population. Conclusions: Approximately 73% of the genotypic variability in nitrate accumulation within the population of RILs arose from differences in nitrate uptake and only 27% from differences in nitrate assimilated, irrespective of whether or not part of the N was recovered as ammonium, or whether the plants were grown in soil or solution culture. Genotypic variability in nitrate accumulation was associated with changes in concentrations of other endogenous solutes (especially carboxylates and soluble carbohydrates) and of tissue water, which minimised differences in osmotic potential of shoot sap between RILs. This offers the opportunity of using the regulation of these solutes as additional traits to manipulate nitrate accumulation. © 2011 Springer Science+Business Media B.V

A novel lung explant model for the ex vivo study of efficacy and mechanisms of anti-influenza drugs

Influenza A virus causes considerable morbidity and mortality largely because of a lack of effective antiviral drugs. Viral neuraminidase inhibitors, which inhibit viral release from the infected cell, are currently the only approved drugs for influenza, but have recently been shown to be less effective than previously thought. Growing resistance to therapies that target viral proteins has led to increased urgency in the search for novel anti-influenza compounds. However, discovery and development of new drugs have been restricted because of differences in susceptibility to influenza between animal models and humans and a lack of translation between cell culture and in vivo measures of efficacy. To circumvent these limitations, we developed an experimental approach based on ex vivo infection of human bronchial tissue explants and optimized a method of flow cytometric analysis to directly quantify infection rates in bronchial epithelial tissues. This allowed testing of the effectiveness of TVB024, a vATPase inhibitor that inhibits viral replication rather than virus release, and to compare efficacy with the current frontline neuraminidase inhibitor, oseltamivir. The study showed that the vATPase inhibitor completely abrogated epithelial cell infection, virus shedding, and the associated induction of proinflammatory mediators, whereas oseltamivir was only partially effective at reducing these mediators and ineffective against innate responses. We propose, therefore, that this explant model could be used to predict the efficacy of novel anti-influenza compounds targeting diverse stages of the viral replication cycle, thereby complementing animal models and facilitating progression of new drugs into clinical trials

Double haploids, markers and QTL analysis in vegetable brassicas

Double haploid (DH) plants of Brassica spp. can be produced via anther culture or culture of microspores. This paper reviews the uses of double haploids in crop improvement research in vegetable brassicas (B. oleracea). Applications of DH lines are described for breeding; construction of linkage maps; genetic analysis of quantitative traits and capturing genetic variation. The advantages and disadvantages of DH lines are discussed

U-BIOPRED clinical adult asthma clusters linked to a subset of sputum omics

Background: Asthma is a heterogeneous disease in which there is a differential response to asthma treatments. This heterogeneity needs to be evaluated so that a personalized management approach can be provided. Objectives: We stratified patients with moderate-to-severe asthma based on clinicophysiologic parameters and performed an omics analysis of sputum. Methods: Partition-around-medoids clustering was applied to a training set of 266 asthmatic participants from the European Unbiased Biomarkers for the Prediction of Respiratory Diseases Outcomes (U-BIOPRED) adult cohort using 8 prespecified clinic-physiologic variables. This was repeated in a separate validation set of 152 asthmatic patients. The clusters were compared based on sputum proteomics and transcriptomics data. Results: Four reproducible and stable clusters of asthmatic patients were identified. The training set cluster T1 consists of patients with well-controlled moderate-to-severe asthma, whereas cluster T2 is a group of patients with late-onset severe asthma with a history of smoking and chronic airflow obstruction. Cluster T3 is similar to cluster T2 in terms of chronic airflow obstruction but is composed of nonsmokers. Cluster T4 is predominantly composed of obese female patients with uncontrolled severe asthma with increased exacerbations but with normal lung function. The validation set exhibited similar clusters, demonstrating reproducibility of the classification. There were significant differences in sputum proteomics and transcriptomics between the clusters. The severe asthma clusters (T2, T3, and T4) had higher sputum eosinophilia than cluster T1, with no differences in sputum neutrophil counts and exhaled nitric oxide and serum IgE levels. Conclusion: Clustering based on clinicophysiologic parameters yielded 4 stable and reproducible clusters that associate with different pathobiological pathways