Covering Chemical Diversity of Genetically-Modified Tomatoes Using Metabolomics for Objective Substantial Equivalence Assessment

As metabolomics can provide a biochemical snapshot of an organism's phenotype it is a promising approach for charting the unintended effects of genetic modification. A critical obstacle for this application is the inherently limited metabolomic coverage of any single analytical platform. We propose using multiple analytical platforms for the direct acquisition of an interpretable data set of estimable chemical diversity. As an example, we report an application of our multi-platform approach that assesses the substantial equivalence of tomatoes over-expressing the taste-modifying protein miraculin. In combination, the chosen platforms detected compounds that represent 86% of the estimated chemical diversity of the metabolites listed in the LycoCyc database. Following a proof-of-safety approach, we show that % had an acceptable range of variation while simultaneously indicating a reproducible transformation-related metabolic signature. We conclude that multi-platform metabolomics is an approach that is both sensitive and robust and that it constitutes a good starting point for characterizing genetically modified organisms

BOTÂNICA E FISIOLOGIA/ BOTANY AND PHYSIOLOGY ANTIOXIDANT ACTIVITY AND PHYSICOCHEMICAL PARAMETERS IN ‘CUERNAVAQUEÑA’ MEXICAN PLUM (Spondias purpurea L.) AT DIFFERENT RIPENING STAGES

ABSTRACT Mexican plum (Spondias purpurea L.) ‘Cuernavaqueña’ was harvested at four ripening stages, with the aim of evaluating the concentration of bioactive compounds and antioxidant capacity in the pulp and the epicarp. The highest ethylene production (9.43 mL kg h-1) and total soluble solids concentration (23.9 °Brix) was observed in the fully ripe stage. Titratable acidity was higher in green stage compared to other analyzed ripening stages in both pulp (0.48 %) and epicarp (0.32 %). Fully ripe plum epicarp presented the highest content of total phenols (GAE 190 mg g-1), flavonoids (QE 214 mg g-1), and carotenoids (853 mg g-1) compared to other ripening stages. The antioxidant capacity was higher in the epicarp of the fully ripe fruit compared to the other ripening stages: DPPH (1087 µM TE /100 g), ABTS (1534 µM TE/100 g), and FRAP (1764 µM TE/100 g). Significant correlations (r = 0.60 *** to 0.95 **) between bioactive compounds concentrations and antioxidant activity were obtained

Indução do amadurecimento de peras 'Rocha' submetidas à baixa temperatura e à aplicação de etileno

O objetivo deste trabalho foi avaliar o efeito da exposição ao frio e da aplicação de etileno, em diferentes períodos, sobre o amadurecimento e a qualidade de peras 'Rocha'. Foram conduzidos dois experimentos. No experimento 1, foram avaliados quatro períodos de exposição a 3ºC (0, 14, 28 e 42 dias) e o efeito da aplicação de 100 ppm de C2H4 por um e dois dias. No experimento 2, foram testados cinco períodos de exposição a 3ºC (0, 15, 30, 45 e 60 dias). No experimento 1, o aumento do período de frio de 14 para 28 dias ocasionou maior amarelecimento da casca e aumento das taxas respiratória e de produção de etileno. Em todos os tratamentos, os frutos apresentaram menor firmeza de polpa e de força para penetração da polpa, em comparação ao controle; no entanto, apresentaram textura adequada para consumo após sete dias em condição ambiente. No experimento 2, a firmeza de polpa, a força para penetração da polpa, a cor verde da casca e a acidez titulável diminuiram com o aumento do período de frio. A exposição dos frutos ao frio de 3ºC, por 15 dias, e a aplicação de 100 ppm de C2H4, por dois dias, são indicadas para induzir o amadurecimento de peras 'Rocha'

Comparative proteomic analysis on fruit ripening processes in two varieties of tropical mango (Mangifera indica)

Mango (Mangifera indica L.) is an economically important fruit. However, the marketability of mango is affected by the perishable nature and short shelf-life of the fruit. Therefore, a better understanding of the mango ripening process is of great importance towards extending its postharvest shelf life. Proteomics is a powerful tool that can be used to elucidate the complex ripening process at the cellular and molecular levels. This study utilized 2-dimensional gel electrophoresis (2D-GE) coupled with MALDI-TOF/TOF to identify differentially abundant proteins during the ripening process of the two varieties of tropical mango, Mangifera indica cv. ‘Chokanan’ and Mangifera indica cv ‘Golden Phoenix’. The comparative analysis between the ripe and unripe stages of mango fruit mesocarp revealed that the differentially abundant proteins identified could be grouped into the three categories namely, ethylene synthesis and aromatic volatiles, cell wall degradation and stress-response proteins. There was an additional category for differential proteins identified from the ‘Chokanan’ variety namely, energy and carbohydrate metabolism. However, of all the differential proteins identified, only methionine gamma-lyase was found in both ‘Chokanan’ and ‘Golden Phoenix’ varieties. Six differential proteins were selected from each variety for validation by analysing their respective transcript expression using reverse transcription-quantitative PCR (RT-qPCR). The results revealed that two genes namely, glutathione S-transferase (GST) and alpha-1,4 glucan phosphorylase (AGP) were found to express in concordant with protein abundant. The findings will provide an insight into the fruit ripening process of different varieties of mango fruits, which is important for postharvest management

On the role of ethylene, auxin and a GOLVEN-like peptide hormone in the regulation of peach ripening

BACKGROUND: In melting flesh peaches, auxin is necessary for system-2 ethylene synthesis and a cross-talk between ethylene and auxin occurs during the ripening process. To elucidate this interaction at the transition from maturation to ripening and the accompanying switch from system-1 to system-2 ethylene biosynthesis, fruits of melting flesh and stony hard genotypes, the latter unable to produce system-2 ethylene because of insufficient amount of auxin at ripening, were treated with auxin, ethylene and with 1-methylcyclopropene (1-MCP), known to block ethylene receptors. The effects of the treatments on the different genotypes were monitored by hormone quantifications and transcription profiling. RESULTS: In melting flesh fruit, 1-MCP responses differed according to the ripening stage. Unexpectedly, 1-MCP induced genes also up-regulated by ripening, ethylene and auxin, as CTG134, similar to GOLVEN (GLV) peptides, and repressed genes also down-regulated by ripening, ethylene and auxin, as CTG85, a calcineurin B-like protein. The nature and transcriptional response of CTG134 led to discover a rise in free auxin in 1-MCP treated fruit. This increase was supported by the induced transcription of CTG475, an IAA-amino acid hydrolase. A melting flesh and a stony hard genotype, differing for their ability to synthetize auxin and ethylene amounts at ripening, were used to study the fine temporal regulation and auxin responsiveness of genes involved in the process. Transcriptional waves showed a tight interdependence between auxin and ethylene actions with the former possibly enhanced by the GLV CTG134. The expression of genes involved in the regulation of ripening, among which are several transcription factors, was similar in the two genotypes or could be rescued by auxin application in the stony hard. Only GLV CTG134 expression could not be rescued by exogenous auxin. CONCLUSIONS: 1-MCP treatment of peach fruit is ineffective in delaying ripening because it stimulates an increase in free auxin. As a consequence, a burst in ethylene production speeding up ripening occurs. Based on a network of gene transcriptional regulations, a model in which appropriate level of CTG134 peptide hormone might be necessary to allow the correct balance between auxin and ethylene for peach ripening to occur is proposed. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12870-016-0730-7) contains supplementary material, which is available to authorized users