Molecular mechanisms of resistance to Myzus persicae conferred by the peach Rm2 gene: A multi-omics view.

The transcriptomic and metabolomic responses of peach to Myzus persicae infestation were studied in Rubira, an accession carrying the major resistance gene Rm2 causing antixenosis, and GF305, a susceptible accession. Transcriptome and metabolome showed both a massive reconfiguration in Rubira 48 hours after infestation while GF305 displayed very limited changes. The Rubira immune system was massively stimulated, with simultaneous activation of genes encoding cell surface receptors involved in pattern-triggered immunity and cytoplasmic NLRs (nucleotide-binding domain, leucine-rich repeat containing proteins) involved in effector-triggered immunity. Hypersensitive reaction featured by necrotic lesions surrounding stylet punctures was supported by the induction of cell death stimulating NLRs/helpers couples, as well as the activation of H2O2-generating metabolic pathways: photorespiratory glyoxylate synthesis and activation of the futile P5C/proline cycle. The triggering of systemic acquired resistance was suggested by the activation of pipecolate pathway and accumulation of this defense hormone together with salicylate. Important reduction in carbon, nitrogen and sulphur metabolic pools and the repression of many genes related to cell division and growth, consistent with reduced apices elongation, suggested a decline in the nutritional value of apices. Finally, the accumulation of caffeic acid conjugates pointed toward their contribution as deterrent and/or toxic compounds in the mechanisms of resistance.

Cuticular waxes of nectarines during fruit development in relation to surface conductance and susceptibility to Monilinia laxa.

The cuticle is composed of cutin and cuticular waxes, and it is the first protective barrier to abiotic and biotic stresses in fruit. In this study, we analysed the composition of and changes in cuticular waxes during fruit development in nectarine (Prunus persica L. Batsch) cultivars, in parallel with their conductance and their susceptibility to Monilinia laxa. The nectarine waxes were composed of triterpenoids, mostly ursolic and oleanolic acids, phytosterols, and very-long-chain aliphatics. In addition, we detected phenolic compounds that were esterified with sugars or with triterpenoids, which are newly described in cuticular waxes. We quantified 42 compounds and found that they changed markedly during fruit development, with an intense accumulation of triterpenoids during initial fruit growth followed by their decrease at the end of endocarp lignification and a final increase in very-long-chain alkanes and hydroxylated triterpenoids until maturity. The surface conductance and susceptibility to Monilinia decreased sharply at the beginning of endocarp lignification, suggesting that triterpenoid deposition could play a major role in regulating fruit permeability and susceptibility to brown rot. Our results provide new insights into the composition of cuticular waxes of nectarines and their changes during fruit development, opening new avenues of research to explore brown rot resistance factors in stone fruit.

A Systems Biology Study in Tomato Fruit Reveals Correlations between the Ascorbate Pool and Genes Involved in Ribosome Biogenesis, Translation, and the Heat-Shock Response.

Changing the balance between ascorbate, monodehydroascorbate, and dehydroascorbate in plant cells by manipulating the activity of enzymes involved in ascorbate synthesis or recycling of oxidized and reduced forms leads to multiple phenotypes. A systems biology approach including network analysis of the transcriptome, proteome and metabolites of RNAi lines for ascorbate oxidase, monodehydroascorbate reductase and galactonolactone dehydrogenase has been carried out in orange fruit pericarp of tomato (Solanum lycopersicum). The transcriptome of the RNAi ascorbate oxidase lines is inversed compared to the monodehydroascorbate reductase and galactonolactone dehydrogenase lines. Differentially expressed genes are involved in ribosome biogenesis and translation. This transcriptome inversion is also seen in response to different stresses in Arabidopsis. The transcriptome response is not well correlated with the proteome which, with the metabolites, are correlated to the activity of the ascorbate redox enzymes-ascorbate oxidase and monodehydroascorbate reductase. Differentially accumulated proteins include metacaspase, protein disulphide isomerase, chaperone DnaK and carbonic anhydrase and the metabolites chlorogenic acid, dehydroascorbate and alanine. The hub genes identified from the network analysis are involved in signaling, the heat-shock response and ribosome biogenesis. The results from this study therefore reveal one or several putative signals from the ascorbate pool which modify the transcriptional response and elements downstream.

Synthesis of new dicinnamoyl 4-deoxy quinic acid and methyl ester derivatives and evaluation of the toxicity against the pea aphid Acyrthosiphon pisum.

New dicinnamoyl (caffeoyl, feruloyl, ortho and para-coumaroyl) 4-deoxyquinic acid and esters were synthesized by using a new 4-deoxy quinic acid triol intermediate. The optimisation of both coupling and deprotection steps allowed the preparation in good yields of the target products either as the carboxylic acid or the methyl ester form. Eight new compounds were evaluated for their ability to influence the feeding behaviour of the pea aphid Acyrthosiphon pisum. Artificial diet bioassays showed that two compounds are toxic (mortality and growth inhibition) at lower concentrations than the reference 3,5-dicaffeoyl quinic acid.

Isolation of C-glycosyl xanthones from Coffea pseudozanguebariae and their location.

The biochemical composition of leaves from Coffea pseudozanguebariae, a wild caffeine-free coffee species, was determined. Two phenolic compounds were extracted from leaves, separated and characterized. Their structures were elucidated by mass spectrometry, and 1D and 2D NMR spectroscopy and were shown to be mangiferin (1) and isomangiferin (2), which were the main polyphenol products. Multiphoton fluorescence imaging was performed to visualize polyphenol distribution in leaf cross sections. Consistent biochemical analysis cell imaging techniques on leaves revealed yellow fluorescence in the epidermis and parenchyma cells corresponding to xanthone compounds.

Volatile organic compound emissions induced by the aphid Myzus persicae differ among resistant and susceptible peach cultivars and a wild relative.

Little is known on aphid-induced emissions of volatile organic compounds (VOCs) from trees and particularly on their intraspecific variability in association with resistance traits. We compared VOC emissions from five peach cultivars (Prunus persica (L.) Batsch) and a wild relative (Prunus davidiana (Carrière) Franch) that differ in their level (susceptible/resistant) and type (antixenosis, antibiosis) of resistance to the green peach aphid, Myzus persicae (Sulzer). Additionally, the kinetics of VOC induction in response to aphids was compared with that by mechanical wounding. Qualitative and overall quantitative differences among peach genotypes were found in VOC emissions that were mainly composed of methyl-salicylate, farnesenes, (E)-ß-ocimene and (E)-4,8-dimethyl-1,3,7-nonatriene. Irrespective of the type of resistance, all resistant genotypes had increased VOC emissions upon aphid attack, while in susceptible genotypes emissions remained low. Emission increases were highest in the genotypes that express increased aphid resistance during second infestations, which had also the highest proportions of methyl-salicylate in their emissions. VOC induction by aphids proceeded slowly with a delay of several hours. Artificial wounding of leaves did not result in emissions of aphid-induced VOCs but caused an immediate burst of green leaf volatiles and benzaldehyde. We conclude that VOC induction in resistant peach cultivars is part of a general defence syndrome that is being avoided or suppressed by M. persicae in the susceptible genotypes. The induction likely involves an aphid-specific elicitor and (methyl)-salicylate in the subsequent signalling and regulation processes that should include gene activation due to the marked delay in the emission response. The results are compared with those of the literature and discussed in view of their ecological and environmental significance.

An integrative genomics approach for deciphering the complex interactions between ascorbate metabolism and fruit growth and composition in tomato.

Very few reports have studied the interactions between ascorbate and fruit metabolism. In order to get insights into the complex relationships between ascorbate biosynthesis/recycling and other metabolic pathways in the fruit, we undertook a fruit systems biology approach. To this end, we have produced tomato transgenic lines altered in ascorbate content and redox ratio by RNAi-targeting several key enzymes involved in ascorbate biosynthesis (2 enzymes) and recycling (2 enzymes). In the VTC (ViTamin C) Fruit project, we then generated phenotypic and genomic (transcriptome, proteome, metabolome) data from wild type and mutant tomato fruit at two stages of fruit development, and developed or implemented statistical and bioinformatic tools as a web application (named VTC Tool box) necessary to store, analyse and integrate experimental data in tomato. By using Kohonen's self-organizing maps (SOMs) to cluster the biological data, pair-wise Pearson correlation analyses and simultaneous visualization of transcript/protein and metabolites (MapMan), this approach allowed us to uncover major relationships between ascorbate and other metabolic pathways.

How does tomato quality (sugar, acid, and nutritional quality) vary with ripening stage, temperature, and irradiance?

The objective of this study was to understand the respective impact of ripening stage, temperature, and irradiance on seasonal variations of tomato fruit quality. During ripening, concentrations in reducing sugars, carotenes, ascorbate, rutin, and caffeic acid derivates increased, whereas those in titratable acidity, chlorophylls, and chlorogenic acid content decreased. Fruit temperature and irradiance affected final fruit composition. Sugars and acids (linked to fruit gustative quality) were not considerably modified, but secondary metabolites with antioxidant properties were very sensitive to fruit environment. Increased fruit irradiance enhanced ascorbate, lycopene, beta-carotene, rutin, and caffeic acid derivate concentrations and the disappearance of oxidized ascorbate and chlorophylls. Increasing the temperature from 21 to 26 degrees C reduced total carotene content without affecting lycopene content. A further temperature increase from 27 to 32 degrees C reduced ascorbate, lycopene, and its precursor's content, but enhanced rutin, caffeic acid derivates, and glucoside contents. The regulation by light and temperature of the biosynthesis pathways of secondary metabolites is discussed.