Rumor Has It…: Relay Communication of Stress Cues in Plants

Recent evidence demonstrates that plants are able not only to perceive and adaptively respond to external information but also to anticipate forthcoming hazards and stresses. Here, we tested the hypothesis that unstressed plants are able to respond to stress cues emitted from their abiotically-stressed neighbors and in turn induce stress responses in additional unstressed plants located further away from the stressed plants. Pisum sativum plants were subjected to drought while neighboring rows of five unstressed plants on both sides, with which they could exchange different cue combinations. On one side, the stressed plant and its unstressed neighbors did not share their rooting volumes (UNSHARED) and thus were limited to shoot communication. On its other side, the stressed plant shared one of its rooting volumes with its nearest unstressed neighbor and all plants shared their rooting volumes with their immediate neighbors (SHARED), allowing both root and shoot communication. Fifteen minutes following drought induction, significant stomatal closure was observed in both the stressed plants and their nearest unstressed SHARED neighbors, and within one hour, all SHARED neighbors closed their stomata. Stomatal closure was not observed in the UNSHARED neighbors. The results demonstrate that unstressed plants are able to perceive and respond to stress cues emitted by the roots of their drought-stressed neighbors and, via ‘relay cuing’, elicit stress responses in further unstressed plants. Further work is underway to study the underlying mechanisms of this new mode of plant communication and its possible adaptive implications for the anticipation of forthcoming abiotic stresses by plants

Not just shrivelling: time-series profiling of the biochemical changes in Corvina (Vitis vinifera L.) berries subjected to post-harvest withering

6openInternationalBothThe grape berry withering process is often seen as a means to concentrate the constituents of the berry via water removal, however, recent molecular studies have indicated that the reprogramming of biosynthetic pathways that impart the unique aroma and flavour of the final wine also occur. Metabolic analysis was performed using GC-MS and LC-MS on Corvina berry dehydrated for up to 108 days. The temporal pattern of metabolic changes and time-series relationship between various classes of metabolites were investigated on berries sampled at 23-time points. Principal component analysis of both GC-MS and LC-MS datasets revealed three distinct phases of post-harvest withering; early (day 0–28), mid (day 35–66) and late (day 69–108) stages. Stress-associated amino acids such as proline, serine, ethanolamine, and leucine accumulated significantly during the mid and late stages of the drying process while phosphorylated glucose and fructose, sucrose, kestose and resveratrol increased massively across time. Unlike most of the identified metabolites, low molecular weight flavanols exhibited a consistent pattern of decline during the withering period. Our network analysis revealed that increased metabolic network connectivity occurred during the middle stage of withering, thus reflecting more coordinated metabolite changes while reduced network connectivity at early and late stages indicates minimal metabolite perturbation during these stages. The current prolonged berry withering experiment revealed the timing of metabolite interconversions in the central metabolism and provided critical clues that link the concentration effect, protein degradation, and the onset of stress-like conditions in drying berriesopenDegu, A.; Wong, D.C.J.; Ciman, G.M.; Lonardi, F.; Mattivi, F.; Fait, A.Degu, A.; Wong, D.C.J.; Ciman, G.M.; Lonardi, F.; Mattivi, F.; Fait, A

Metabolic and physiological responses of shiraz and cabernet sauvignon (Vitis vinifera L.) to near optimal temperatures of 25 and 35 \ub0C

Shiraz and Cabernet Sauvignon (Cs) grapevines were grown at near optimal temperatures (25 or 35 \ub0C). Gas exchange, fluorescence, metabolic profiling and correlation based network analysis were used to characterize leaf physiology. When grown at 25 \ub0C, the growth rate and photosynthesis of both cultivars were similar. At 35 \ub0C Shiraz showed increased respiration, non-photochemical quenching and reductions of photosynthesis and growth. In contrast, Cs maintained relatively stable photosynthetic activity and growth regardless of the condition. In both cultivars, growth at 35 \ub0C resulted in accumulations of secondary sugars (raffinose, fucose and ribulose) and reduction of primary sugars concentration (glucose, fructose and sucrose), more noticeably in Shiraz than Cs. In spite of similar patterns of metabolic changes in response to growth at 35 \ub0C, significant differences in important leaf antioxidants and antioxidant precursors (DHA/ascorbate, quinates, cathechins) characterized the cultivar response. Correlation analysis reinforced Shiraz sensitivity to the 35 \ub0C, showing higher number of newly formed edges at 35 \ub0C and higher modularity in Shiraz as compared to Cs. The results suggest that the optimal growth temperatures of grapevines are cultivar dependent, and allow a first insight into the variability of the metabolic responses of grapevines under varied temperatures

Conserved changes in dynamics of metabolic processes during fruit development and ripening across species

Computational analyses of molecular phenotypes traditionally aim at identifying biochemical components that exhibit differential expression under various scenarios (e.g. environmental and internal perturbations) in a single species. High-throughput metabolomics technologies allow the quantification of (relative) metabolite levels across developmental stages in different tissues, organs, and species. Novel methods for analyzing the resulting multiple data tables could reveal preserved dynamics of metabolic processes across species. The problem we address in this study is 2-fold. (1) We derive a single data table, referred to as a compromise, which captures information common to the investigated set of multiple tables containing data on different fruit development and ripening stages in three climacteric (i.e. peach [Prunus persica] and two tomato [Solanum lycopersicum] cultivars, Ailsa Craig and M82) and two nonclimacteric (i.e. strawberry [Fragaria × ananassa] and pepper [Capsicum chilense]) fruits; in addition, we demonstrate the power of the method to discern similarities and differences between multiple tables by analyzing publicly available metabolomics data from three tomato ripening mutants together with two tomato cultivars. (2) We identify the conserved dynamics of metabolic processes, reflected in the data profiles of the corresponding metabolites that contribute most to the determined compromise. Our analysis is based on an extension to principal component analysis, called STATIS, in combination with pathway overenrichment analysis. Based on publicly available metabolic profiles for the investigated species, we demonstrate that STATIS can be used to identify the metabolic processes whose behavior is similarly affected during fruit development and ripening. These findings ultimately provide insights into the pathways that are essential during fruit development and ripening across species.Fil: Klie, Sebastian. Max Planck Institute of Molecular Plant Physiology; AlemaniaFil: Osorio, Sonia. Consejo Superior de Investigaciones Cientificas. Instituto de Hortofruticultura Subtropical y Mediterránea; EspañaFil: Tohge, Takayuki. Max Planck Institute of Molecular Plant Physiology; AlemaniaFil: Drincovich, Maria Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Rosario. Centro de Estudios Fotosintéticos y Bioquímicos (i); ArgentinaFil: Fait, Aaron. Ben-Gurion University of the Negrev; IsraelFil: Giovannoni, Federico. Cornell University; Estados UnidosFil: Fernie, Alisdair R.. Max Planck Institute of Molecular Plant Physiology; AlemaniaFil: Nikoloski, Zoran. Max Planck Institute of Molecular Plant Physiology; Alemani

Transcriptome and metabolite profiling reveals that prolonged drought modulates the phenylpropanoid and terpenoid pathway in white grapes (Vitis vinifera L.)

Background: Secondary metabolism contributes to the adaptation of a plant to its environment. In wine grapes, fruit secondary metabolism largely determines wine quality. Climate change is predicted to exacerbate drought events in several viticultural areas, potentially affecting the wine quality. In red grapes, water deficit modulates flavonoid accumulation, leading to major quantitative and compositional changes in the profile of the anthocyanin pigments; in white grapes, the effect of water deficit on secondary metabolism is still largely unknown Results: In this study we investigated the impact of water deficit on the secondary metabolism of white grapes using a large scale metabolite and transcript profiling approach in a season characterized by prolonged drought. Irrigated grapevines were compared to non-irrigated grapevines that suffered from water deficit from early stages of berry development to harvest. A large effect of water deficit on fruit secondary metabolism was observed. Increased concentrations of phenylpropanoids, monoterpenes, and tocopherols were detected, while carotenoid and flavonoid accumulations were differentially modulated by water deficit according to the berry developmental stage. The RNA- sequencing analysis carried out on berries collected at three developmental stages—before, at the onset, and at late ripening—indicated that water deficit affected the expression of 4,889 genes. The Gene Ontology category secondary metabolic process was overrepresented within up-regulated genes at all the stages of fruit development considered, and within down-regulated genes before ripening. Eighteen phenylpropanoid, 16 flavonoid, 9 carotenoid, and 16 terpenoid structural genes were modulated by water deficit, indicating the transcriptional regulation of these metabolic pathways in fruit exposed to water deficit. An integrated network and promoter analyses identified a transcriptional regulatory module that encompasses terpenoid genes, transcription factors, and enriched drought- responsive elements in the promoter regions of those genes as part of the grapes response to drought Conclusion: Our study reveals that grapevine berries respond to drought by modulating several secondary metabolic pathways, and particularly, by stimulating the production of phenylpropanoids, the carotenoid zeaxanthin, and of volatile organic compounds such as monoterpenes, with potential effects on grape and wine antioxidant potential, composition, and sensory feature

Rootstock identity in melon-pumpkin graft combinations determines fruit metabolite profile

Grafting has the potential to improve melon fruit yield and quality, but it is currently held that a lack of compatibility between the rootstock and scion compromises such an effect. To throw light on this subject, we studied melon-pumpkin graft combinations with different levels of compatibility to assess to the effect of the rootstock identity on melon fruit yield and quality, including total fruit ortho-diphenols, total flavonoids, and primary fruit metabolites. Melon cv. ‘Kiran’ (Ki) was grafted onto three pumpkin rootstocks, ‘TZ-148’ (TZ), ‘Shimshon’ (Sh), and ‘53006’ (r53), characterized by high, moderate, and low compatibility, respectively. The non-grafted melon cultivar Ki was used as the control. The incompatible combination Ki/r53 gave the lowest fruit yield and the lowest average fruit weight. In that combination, the content of total ortho-diphenols increased vs. Ki and Ki/TZ and that of total flavonoids decreased vs. Ki/Sh. In addition, concentrations of the amino acids, glutamate, methionine, valine, alanine, glycine, and serine, increased in the pulp of the two compatible combinations, i.e., Ki/TZ and Ki/Sh, suggesting that rootstock identity and compatibility with melon Ki scion modulated amino acid synthesis. Our results show an association between rootstock identity (and level of compatibility with the scion) and an enhancement of fruit nutritional values, i.e., high concentrations of organic acids (determined as citrate, malate, fumarate, and succinate) and soluble carbohydrates (sucrose) were recorded in the pulp of the two compatible combinations, i.e., Ki/TZ and Ki/Sh

Cytoplasmic genome substitution in wheat affects the nuclear-cytoplasmic cross-talk leading to transcript and metabolite alterations

Abstract Background Alloplasmic lines provide a unique tool to study nuclear-cytoplasmic interactions. Three alloplasmic lines, with nuclear genomes from Triticum aestivum and harboring cytoplasm from Aegilops uniaristata, Aegilops tauschii and Hordeum chilense, were investigated by transcript and metabolite profiling to identify the effects of cytoplasmic substitution on nuclear-cytoplasmic signaling mechanisms. Results In combining the wheat nuclear genome with a cytoplasm of H. chilense, 540 genes were significantly altered, whereas 11 and 28 genes were significantly changed in the alloplasmic lines carrying the cytoplasm of Ae. uniaristata or Ae. tauschii, respectively. We identified the RNA maturation-related process as one of the most sensitive to a perturbation of the nuclear-cytoplasmic interaction. Several key components of the ROS chloroplast retrograde signaling, together with the up-regulation of the ROS scavenging system, showed that changes in the chloroplast genome have a direct impact on nuclear-cytoplasmic cross-talk. Remarkably, the H. chilense alloplasmic line down-regulated some genes involved in the determination of cytoplasmic male sterility without expressing the male sterility phenotype. Metabolic profiling showed a comparable response of the central metabolism of the alloplasmic and euplasmic lines to light, while exposing larger metabolite alterations in the H. chilense alloplasmic line as compared with the Aegilops lines, in agreement with the transcriptomic data. Several stress-related metabolites, remarkably raffinose, were altered in content in the H. chilense alloplasmic line when exposed to high light, while amino acids, as well as organic acids were significantly decreased. Alterations in the levels of transcript, related to raffinose, and the photorespiration-related metabolisms were associated with changes in the level of related metabolites. Conclusion The replacement of a wheat cytoplasm with the cytoplasm of a related species affects the nuclear-cytoplasmic cross-talk leading to transcript and metabolite alterations. The extent of these modifications was limited in the alloplasmic lines with Aegilops cytoplasm, and more evident in the alloplasmic line with H. chilense cytoplasm. We consider that, this finding might be linked to the phylogenetic distance of the genomes.The authors thank Prof. S.S. Maan (North Dakota State University, USA) for providing the alloplasmic lines T183 and T195 and Dr. Alessio Aprile (University of Salento, Lecce, Italy) for help in data analysis. LQ was partly supported by grant # 2770367, the Chief Scientist Fund, the Ministry of Agriculture, Israel.Peer Reviewe

Multi-Omics and Integrated Network Analyses Reveal New Insights into the Systems Relationships between Metabolites, Structural Genes, and Transcriptional Regulators in Developing Grape Berries (Vitis vinifera L.) Exposed to Water Deficit

Grapes are one of the major fruit crops and they are cultivated in many dry environments. This study comprehensively characterizes the metabolic response of grape berries exposed to water deficit at different developmental stages. Increases of proline, branched-chain amino acids, phenylpropanoids, anthocyanins, and free volatile organic compounds have been previously observed in grape berries exposed to water deficit. Integrating RNA-sequencing analysis of the transcriptome with large-scale analysis of central and specialized metabolites, we reveal that these increases occur via a coordinated regulation of key structural pathway genes. Water deficit-induced up-regulation of flavonoid genes is also coordinated with the down-regulation of many stilbene synthases and a consistent decrease in stilbenoid concentration. Water deficit activated both ABA-dependent and ABA-independent signal transduction pathways by modulating the expression of several transcription factors. Gene-gene and gene-metabolite network analyses showed that water deficit-responsive transcription factors such as bZIPs, AP2/ERFs, MYBs, and NACs are implicated in the regulation of stress-responsive metabolites. Enrichment of known and novel cis-regulatory elements in the promoters of several ripening-specific/water deficit-induced modules further affirms the involvement of a transcription factor cross-talk in the berry response to water deficit. Together, our integrated approaches show that water deficit-regulated gene modules are strongly linked to key fruit-quality metabolites and multiple signal transduction pathways may be critical to achieve a balance between the regulation of the stress-response and the berry ripening program. This study constitutes an invaluable resource for future discoveries and comparative studies, in grapes and other fruits, centered on reproductive tissue metabolism under abiotic stress.This study was funded by the European Territorial Cooperation program (Sustainable viticulture and improvement of the territorial resources of the grape and wine industry), the Fondazione Edmund Mach (GMPF Program), the COST Action FA1106 Quality Fruit, Genome British Columbia (10R21188), and the Natural Sciences and Engineering Research Council of Canada (10R23082)

Metabolic Profiling of a Mapping Population Exposes New Insights in the Regulation of Seed Metabolism and Seed, Fruit, and Plant Relations

To investigate the regulation of seed metabolism and to estimate the degree of metabolic natural variability, metabolite profiling and network analysis were applied to a collection of 76 different homozygous tomato introgression lines (ILs) grown in the field in two consecutive harvest seasons. Factorial ANOVA confirmed the presence of 30 metabolite quantitative trait loci (mQTL). Amino acid contents displayed a high degree of variability across the population, with similar patterns across the two seasons, while sugars exhibited significant seasonal fluctuations. Upon integration of data for tomato pericarp metabolite profiling, factorial ANOVA identified the main factor for metabolic polymorphism to be the genotypic background rather than the environment or the tissue. Analysis of the coefficient of variance indicated greater phenotypic plasticity in the ILs than in the M82 tomato cultivar. Broad-sense estimate of heritability suggested that the mode of inheritance of metabolite traits in the seed differed from that in the fruit. Correlation-based metabolic network analysis comparing metabolite data for the seed with that for the pericarp showed that the seed network displayed tighter interdependence of metabolic processes than the fruit. Amino acids in the seed metabolic network were shown to play a central hub-like role in the topology of the network, maintaining high interactions with other metabolite categories, i.e., sugars and organic acids. Network analysis identified six exceptionally highly co-regulated amino acids, Gly, Ser, Thr, Ile, Val, and Pro. The strong interdependence of this group was confirmed by the mQTL mapping. Taken together these results (i) reflect the extensive redundancy of the regulation underlying seed metabolism, (ii) demonstrate the tight co-ordination of seed metabolism with respect to fruit metabolism, and (iii) emphasize the centrality of the amino acid module in the seed metabolic network. Finally, the study highlights the added value of integrating metabolic network analysis with mQTL mapping