Luca Tardella

Use of network analysis to capture key traits affecting tomato organoleptic quality

The long-term objective of tomato breeders is to identify metabolites that contribute to defining the target flavour and to design strategies to enhance it. This paper reports the results of network analysis, based on metabolic phenotypic and sensory data, to highlight important relationships among such traits. This tool allowed a reduction in data set complexity, building a network consisting of 35 nodes and 74 links corresponding to the 74 significant (positive or negative) correlations among the variables studied. A number of links among traits contributing to fruit organoleptic quality and to the perception of sensory attributes were identified. Modular partitioning of the characteristics involved in fruit organoleptic perception captured the essential fruit parameters that regulate interactions among different class traits. The main feature of the network was the presence of three nodes interconnected among themselves (dry matter, pH, and °Brix) and with other traits, and nodes with widely different linkage degrees. Identification of strong associations between some metabolic and sensory traits, such as citric acid with tomato smell, glycine with tomato smell, and granulosity with dry matter, suggests a basis for more targeted investigations in the future.

Unraveling the complexity of transcriptomic, metabolomic and quality environmental response of tomato fruit

BackgroundThe environment has a profound influence on the organoleptic quality of tomato (Solanum lycopersicum) fruit, the extent of which depends on a well-regulated and dynamic interplay among genes, metabolites and sensorial attributes. We used a systems biology approach to elucidate the complex interacting mechanisms regulating the plasticity of sensorial traits. To investigate environmentally challenged transcriptomic and metabolomic remodeling and evaluate the organoleptic consequences of such variations we grown three tomato varieties, Heinz 1706, whose genome was sequenced as reference and two “local” ones, San Marzano and Vesuviano in two different locations of Campania region (Italy).ResultsResponses to environment were more pronounced in the two “local” genotypes, rather than in the Heinz 1706. The overall genetic composition of each genotype, acting in trans, modulated the specific response to environment. Duplicated genes and transcription factors, establishing different number of network connections by gaining or losing links, play a dominant role in shaping organoleptic profile. The fundamental role of cell wall metabolism in tuning all the quality attributes, including the sensorial perception, was also highlighted.ConclusionsAlthough similar fruit-related quality processes are activated in the same environment, different tomato genotypes follow distinct transcriptomic, metabolomic and sensorial trajectories depending on their own genetic makeup.

Identification of Relevant Conformational Epitopes on the HER2 Oncoprotein by Using Large Fragment Phage Display (LFPD)

We developed a new phage-display based approach, the Large Fragment Phage Display (LFPD), that can be used for mapping conformational epitopes on target molecules of immunological interest. LFPD uses a simplified and more effective phage-display approach in which only a limited set of larger fragments (about 100 aa in length) are expressed on the phage surface. Using the human HER2 oncoprotein as a target, we identified novel B-cell conformational epitopes. The same homologous epitopes were also detected in rat HER2 and all corresponded to the epitopes predicted by computational analysis (PEPITO software), showing that LFPD gives reproducible and accurate results. Interestingly, these newly identified HER2 epitopes seem to be crucial for an effective immune response against HER2-overexpressing breast cancers and might help discriminating between metastatic breast cancer and early breast cancer patients. Overall, the results obtained in this study demonstrated the utility of LFPD and its potential application to the detection of conformational epitopes on many other molecules of interest, as well as, the development of new and potentially more effective B-cell conformational epitopes based vaccines.

Identifiability and inferential issues in capture-recapture experiments with heterogeneous detection probabilities

We focus on a capture-recapture model in which capture prob- abilities arise from an unspecified distribution F. We show that model pa- rameters are identifiable based on the unconditional likelihood. This is not true with the conditional likelihood. We also clarify that consistency and asymptotic equivalence of maximum likelihood estimators based on condi- tional and unconditional likelihood do not hold. We show that estimates of the undetected fraction of population based on the unconditional likelihood converge to the so-called estimable sharpest lower bound and we derive a new asymptotic equivalence result. We finally provide theoretical and sim- ulation arguments in favor of the use of the unconditional likelihood rather than the conditional likelihood especially when one is willing to infer on the sharpest lower bound. AMS 2000 subject classifications: Primary 62G10; secondary 62F12.

Improved inference on capture recapture models with behavioural effects

In the context of capture-recapture modeling for estimating the unknown size of a finite population it is often required a flexible framework for dealing with a behavioural response to trapping. Many alternative settings have been proposed in the literature to account for the variation of capture probability at each occasion depending on the previous capture history. Inference is typically carried out relying on the so-called conditional likelihood approach. We highlight that such approach may, with positive probability, lead to inferential pathologies such as unbounded estimates for the finite size of the population. The occurrence of such likelihood failures is characterized within a very general class of behavioural effect models. It is also pointed out that a fully Bayesian analysis overcomes the likelihood failure phenomenon. The overall improved performance of alternative Bayesian estimators is investigated under different non-informative prior distributions verifying their comparative merits with both simulated and real data.

Improved Harmonic Mean Estimator for Phylogenetic Model Evidence

Bayesian phylogenetic methods are generating noticeable enthusiasm in the field of molecular systematics. Many phylogenetic models are often at stake, and different approaches are used to compare them within a Bayesian framework. The Bayes factor, defined as the ratio of the marginal likelihoods of two competing models, plays a key role in Bayesian model selection. We focus on an alternative estimator of the marginal likelihood whose computation is still a challenging problem. Several computational solutions have been proposed, none of which can be considered outperforming the others simultaneously in terms of simplicity of implementation, computational burden and precision of the estimates. Practitioners and researchers, often led by available software, have privileged so far the simplicity of the harmonic mean (HM) estimator. However, it is known that the resulting estimates of the Bayesian evidence in favor of one model are biased and often inaccurate, up to having an infinite variance so that the reliability of the corresponding conclusions is doubtful. We consider possible improvements of the generalized harmonic mean (GHM) idea that recycle Markov Chain Monte Carlo (MCMC) simulations from the posterior, share the computational simplicity of the original HM estimator, but, unlike it, overcome the infinite variance issue. We show reliability and comparative performance of the improved harmonic mean estimators comparing them to approximation techniques relying on improved variants of the thermodynamic integration.

Bayesian Plackett–Luce Mixture Models for Partially Ranked Data

The elicitation of an ordinal judgment on multiple alternatives is often required in many psychological and behavioral experiments to investigate preference/choice orientation of a specific population. The Plackett–Luce model is one of the most popular and frequently applied parametric distributions to analyze rankings of a finite set of items. The present work introduces a Bayesian finite mixture of Plackett–Luce models to account for unobserved sample heterogeneity of partially ranked data. We describe an efficient way to incorporate the latent group structure in the data augmentation approach and the derivation of existing maximum likelihood procedures as special instances of the proposed Bayesian method. Inference can be conducted with the combination of the Expectation-Maximization algorithm for maximum a posteriori estimation and the Gibbs sampling iterative procedure. We additionally investigate several Bayesian criteria for selecting the optimal mixture configuration and describe diagnostic tools for assessing the fitness of ranking distributions conditionally and unconditionally on the number of ranked items. The utility of the novel Bayesian parametric Plackett–Luce mixture for characterizing sample heterogeneity is illustrated with several applications to simulated and real preference ranked data. We compare our method with the frequentist approach and a Bayesian nonparametric mixture model both assuming the Plackett–Luce model as a mixture component. Our analysis on real datasets reveals the importance of an accurate diagnostic check for an appropriate in-depth understanding of the heterogenous nature of the partial ranking data.

Fusarium oxysporum f.sp. radicis-lycopersici induces distinct transcriptome reprogramming in resistant and susceptible isogenic tomato lines

BackgroundFusarium oxysporum f.sp. radicis-lycopersici (FORL) is one of the most destructive necrotrophic pathogens affecting tomato crops, causing considerable field and greenhouse yield losses. Despite such major economic impact, little is known about the molecular mechanisms regulating Fusarium oxysporum f.sp. radicis-lycopersici resistance in tomato.ResultsA transcriptomic experiment was carried out in order to investigate the main mechanisms of FORL response in resistant and susceptible isogenic tomato lines. Microarray analysis at 15 DPI (days post inoculum) revealed a distinct gene expression pattern between the two genotypes in the inoculated vs non-inoculated conditions. A model of plant response both for compatible and incompatible reactions was proposed. In particular, in the incompatible interaction an activation of defense genes related to secondary metabolite production and tryptophan metabolism was observed. Moreover, maintenance of the cell osmotic potential after the FORL challenging was mediated by a dehydration-induced protein. As for the compatible interaction, activation of an oxidative burst mediated by peroxidases and a cytochrome monooxygenase induced cell degeneration and necrosis.ConclusionsOur work allowed comprehensive understanding of the molecular basis of the tomato-FORL interaction. The result obtained emphasizes a different transcriptional reaction between the resistant and the susceptible genotype to the FORL challenge. Our findings could lead to the improvement in disease control strategies.

Tomato Genome-Wide Transcriptional Responses to Fusarium Wilt and Tomato Mosaic Virus

Since gene expression approaches constitute a starting point for investigating plant–pathogen systems, we performed a transcriptional analysis to identify a set of genes of interest in tomato plants infected with F. oxysporum f. sp. lycopersici (Fol) and Tomato Mosaic Virus (ToMV). Differentially expressed tomato genes upon inoculation with Fol and ToMV were identified at two days post-inoculation. A large overlap was found in differentially expressed genes throughout the two incompatible interactions. However, Gene Ontology enrichment analysis evidenced specific categories in both interactions. Response to ToMV seems more multifaceted, since more than 70 specific categories were enriched versus the 30 detected in Fol interaction. In particular, the virus stimulated the production of an invertase enzyme that is able to redirect the flux of carbohydrates, whereas Fol induced a homeostatic response to prevent the fungus from killing cells. Genomic mapping of transcripts suggested that specific genomic regions are involved in resistance response to pathogen. Coordinated machinery could play an important role in prompting the response, since 60% of pathogen receptor genes (NB-ARC-LRR, RLP, RLK) were differentially regulated during both interactions. Assessment of genomic gene expression patterns could help in building up models of mediated resistance responses.

Robust semiparametric mixing for detecting differentially expressed genes in microarray experiments

An important goal of microarray studies is the detection of genes that show significant changes in observed expressions when two or more classes of biological samples such as treatment and control are compared. Using the c-fold rule, a gene is declared to be differentially expressed if its average expression level varies by more than a constant factor c between treatment and control (typically c=2). While often used, however, this simple rule is not completely convincing. By modeling this filter, a binary variable is defined at the genexexperiment level, allowing for a more powerful treatment of the corresponding information. A gene-specific random term is introduced to control for both dependence among genes and variability with respect to the c-fold threshold. Inference is carried out via a two-level finite mixture model under a likelihood approach. Then, parameter estimates are also derived using the counting distribution under a Bayesian nonparametric approach which allows to keep under control some error rate of erroneous discoveries. The effectiveness of both proposed approaches is illustrated through a large-scale simulation study and a well known benchmark data set.