Genotypes of Eruca vesicaria subsp. sativa grown in contrasting field environments differ on transcriptomic and metabolomic levels, significantly impacting nutritional quality.

Rocket (Eruca vesicaria subsp. sativa) is a source of sulfur-containing glucosinolates (GSLs). GSLs and their breakdown hydrolysis products (GHPs) are responsible for health-related benefits, such as anti-cancer and anti-neurodegenerative properties. Understanding how phytochemical composition changes between cultivation environments is key to developing cultivars with improved nutritional quality. Two consecutive harvests (first and second regrowth) of crops, grown in both Italy and the UK, were used to determine the phytochemical and transcriptomic differences between six lines of Eruca. Samples were taken upon delivery from field sites (D0) and after five days of cold storage (D5) for each location. Leaves were analysed for sulfur content, volatile organic compounds (VOCs), GSLs, GHPs, and sugars. Transcriptome data were associated with metabolite profiles to identify differentially expressed genes between plants grown in the two environments. VOC compounds (carbon disulfide, methyl thiocyanate) were associated with growth environment and with differences in sulfur metabolism gene expression (APR2, LSU2, LSU3, SDI1, SiR), GSL biosynthesis (MYB28, FMOGS-OX2) and GHP formation (ESM1, TGG1, TGG2). The concentrations of sugars were an order of magnitude greater in UK grown samples (up to 29.9 mg g-1 dry weight; dw). Sulfur content was significantly higher in the Italy plant samples (11.4 - 20.1 mg g-1 dw), which was in turn associated with higher concentrations of GSLs (pentyl GSL, up to 15.8 µmol g-1 dw; sinigrin, up to 0.005 µmol g-1 dw; glucoraphanin, up to 5.1 µmol g-1 dw; glucorucolamine, up to 23.6 µmol g-1 dw; neoglucobrassicin, up to 5.3 µmol g-1 dw) and hydrolysis products (sativin, up to 13.5 µmol g-1 dw; erucin, up to 1 µmol g-1 dw; sulforaphane, up to 34.7 µmol g-1 dw). VOC profiles of plants cultivated in the UK were distinct from Italy grown plants, with higher relative abundances of alkanes and esters in second cut and shelf-life (D5) samples. The data indicate a significant interaction of cultivar response with environment, highlighting the difficulty of producing Eruca crops with consistent phytochemical and postharvest traits. Genes with differential expression between plants grown in Italy and the UK could be used as markers of phytochemical quality and composition.

Nematically Templated Vortex Lattices in Superconducting FeSe.

New pathways to controlling the morphology of superconducting vortex lattices─and their subsequent dynamics─are required to guide and scale vortex world-lines into a computing platform. We have found that the nematic twin boundaries align superconducting vortices in the adjacent terraces due to the incommensurate potential between vortices surrounding twin boundaries and those trapped within them. With the varying density and morphology of twin boundaries, the vortex lattice assumes several distinct structural phases, including square, regular, and irregular one-dimensional lattices. Through concomitant analysis of vortex lattice models, we have inferred the characteristic energetics of the twin boundary potential and furthermore predicted the existence of geometric size effects as a function of increasing confinement by the twin boundaries. These findings extend the ideas of directed control over vortex lattices to intrinsic topological defects and their self-organized networks, which have direct implications for the future design and control of strain-based topological quantum computing architectures.

Mineral concentrations in milk from cows fed seaweed (Saccharina latissima) under different basal protein supplementation.

Sixteen multiparous Holstein cows in four blocks of 4 × 4 Latin square over 4-week experimental periods were used to study the effects of seaweed (Saccharina latissima) supplement (with/without) and protein source (rapeseed meal (RSM)/wheat distiller's grain (WDG)) on milk mineral concentrations. Dietary treatments did not affect milk production and basic composition. Feeding seaweed slightly decreased milk Ca and Cu concentrations; whilst increased (by 3.3-fold) milk iodine (I) concentration, due to a higher dietary I supply. Substitution of WDG with RSM increased feed-to-milk transfer of Ca, Na, and Se and decreased that of Mg, P, Fe, and Mn; but only reduced milk Mn and I concentrations (the latter by 27 % as a potential result of increased glucosinolate intake). Seaweed supplement can improve milk I content when cows' I supply/availability is limited, but care should be taken to avoid excess milk I contents that may pose nutritional risks for young children.

In vitro application of Eruca vesicaria subsp. sativa leaf extracts and associated metabolites reduces the growth of Oomycota species involved in Kiwifruit Vine Decline Syndrome.

This study aimed to determine whether leaf extracts from seven Eruca vesicaria subsp. sativa cultivars and their biochemically active compounds (glucosinolates and downstream-derived products) inhibit mycelia growth of three well-known pathogenic oomycetes, Phytopythium chamaehyphon, Phytopythium vexans and Phytophthora citrophthora; being the most significant in the development of Kiwifruit Vine Decline Syndrome (KVDS). Leaf extract quantity of 10, 20 and 30 mg were inoculated in Petri dish (90 mm Ø, each 22 mL of liquid medium - Potato Dextrose Agar), for in vitro bioassays. A pathogen plug was placed in the centre of each plate and the Oomycota colony perimeter was marked 5 days after inoculation. Radial colony growth was measured from 4 marks per plate 5, 10, and 15 days after inoculation, further elaborated with Image J software image analysis. Growth rates for all strains were inhibited by around 67% after 15 days. This was most pronounced when applying the highest concentration of leaf extract. By using Liquid Chromatography Mass Spectrometry (LC-MS) and Gas Chromatography Mass Spectrometry (GC-MS), fifteen glucosinolate compounds, of which glucosativin was found in the highest quantity, were identified. Concentrations of hydrolysis products produced by leaves (erucin and sativin) were also investigated, and were significantly associated with colony radial growth, especially towards Pp. chamaehyphon and Pp. vexans. Three downstream products of glucosinolates (two pure isothiocyanates, AITC and PEITC; and one indole I3C; all commonly present in Brassicaceae) were also tested, and a statistically significant inhibition of growth was observed at the highest concentration (0.6 µL).

Concurrent anthropogenic air pollutants enhance recruitment of a specialist parasitoid.

Air pollutants-such as nitrogen oxides, emitted in diesel exhaust, and ozone (O3)-disrupt interactions between plants, the insect herbivore pests that feed upon them and natural enemies of those herbivores (e.g. parasitoids). Using eight field-based rings that emit regulated quantities of diesel exhaust and O3, we investigated how both pollutants, individually and in combination, altered the attraction and parasitism rate of a specialist parasitoid (Diaeretiella rapae) on aphid-infested and un-infested Brassica napus plants. Individual effects of O3 decreased D. rapae abundance and emergence by 37% and 55%, respectively, compared with ambient (control) conditions. When O3 and diesel exhaust were emitted concomitantly, D. rapae abundance and emergence increased by 79% and 181%, respectively, relative to control conditions. This attraction response occurred regardless of whether plants were infested with aphids and was associated with an increase in the concentration of aliphatic glucosinolates, especially gluconapin (3-butenyl-glucosinolate), within B. napus leaves. Plant defensive responses and their ability to attract natural aphid enemies may be beneficially impacted by pollution exposure. These results demonstrate the importance of incorporating multiple air pollutants when considering the effects of air pollution on plant-insect interactions.

Corrigendum: The Eruca sativa genome and transcriptome: A targeted analysis of sulfur metabolism and glucosinolate biosynthesis pre and postharvest.

[This corrects the article DOI: 10.3389/fpls.2020.525102.].

Quantitative trait loci analysis of glucosinolate, sugar, and organic acid concentrations in Eruca vesicaria subsp. sativa.

Eruca vesicaria subsp. sativa is a leafy vegetable of the Brassicaceae family known for its pungency. Variation in growing conditions, leaf age, agronomic practices, and variety choice lead to inconsistent quality, especially in content of isothiocyanates (ITCs) and their precursor glucosinolates (GSLs). We present the first linkage and Quantitative Trait Loci (QTL) map for Eruca, generated using a population of 139 F4 lines. A significant environmental effect on the abundance of primary and secondary metabolites was observed, with UK-grown plants containing significantly higher concentrations of glucoraphanin, malic acid, and total sugars. Italian-grown plants were characterized by higher concentrations of glucoerucin, indolic GSLs, and low monosaccharides. 20 QTL were identified and associated with robust SNP markers. Five genes putatively associated with the synthesis of the GSL 4-methoxyglucobrassicin (4MGB) were identified as candidate regulators underlying QTL. Analysis revealed that orthologs of MYB51, IGMT1 and IGMT4 present on LG1 are associated with 4MGB concentrations in Eruca. This research illustrates the utility of the map for identifying genes associated with nutritional composition in Eruca and its value as a genetic resource to assist breeding programs for this leafy vegetable crop.

Important Odorants of Four Brassicaceae Species, and Discrepancies between Glucosinolate Profiles and Observed Hydrolysis Products.

It is widely accepted that the distinctive aroma and flavour traits of Brassicaceae crops are produced by glucosinolate (GSL) hydrolysis products (GHPs) with other non-GSL derived compounds also reported to contribute significantly to their aromas. This study investigated the flavour profile and glucosinolate content of four Brassicaceae species (salad rocket, horseradish, wasabi, and watercress). Solid-phase microextraction followed by gas chromatography-mass spectrometry and gas chromatography-olfactometry were used to determine the volatile compounds and odorants present in the four species. Liquid chromatography-mass spectrometry was used to determine the glucosinolate composition, respectively. A total of 113 compounds and 107 odour-active components were identified in the headspace of the four species. Of the compounds identified, 19 are newly reported for 'salad' rocket, 26 for watercress, 30 for wasabi, and 38 for horseradish, marking a significant step forward in understanding and characterising aroma generation in these species. There were several non-glucosinolate derived compounds contributing to the 'pungent' aroma profile of the species, indicating that the glucosinolate-derived compounds are not the only source of these sensations in Brassicaceae species. Several discrepancies between observed glucosinolates and hydrolysis products were observed, and we discuss the implications of this for future studies.

Corrigendum: Morphophysiology of Potato (Solanum tuberosum) in Response to Drought Stress: Paving the Way Forward.

[This corrects the article DOI: 10.3389/fpls.2020.597554.].

High Glucosinolate Content in Rocket Leaves (Diplotaxis tenuifolia and Eruca sativa) after Multiple Harvests Is Associated with Increased Bitterness, Pungency, and Reduced Consumer Liking.

Rocket (Diplotaxis tenuifolia and Eruca sativa) leaves delivered to the UK market are variable in appearance, taste, and flavour over the growing season. This study presents sensory and consumer analyses of rocket produce delivered to the UK over the course of one year, and evaluated the contribution of environmental and cultivation factors upon quality traits and phytochemicals called glucosinolates (GSLs). GSL abundance was positively correlated with higher average growth temperatures during the crop cycle, and perceptions of pepperiness, bitterness, and hotness. This in turn was associated with reduced liking, and corresponded to low consumer acceptance. Conversely, leaves with greater sugar content were perceived as more sweet, and had a higher correlation with consumer acceptance of the test panel. First cut leaves of rocket were favoured more by consumers, with multiple leaf cuts associated with low acceptance and higher glucosinolate concentrations. Our data suggest that the practice of harvesting rocket crops multiple times reduces consumer acceptability due to increases in GSLs, and the associated bitter, hot, and peppery perceptions some of their hydrolysis products produce. This may have significant implications for cultivation practices during seasonal transitions, where leaves typically receive multiple harvests and longer growth cycles.

The Eruca sativa Genome and Transcriptome: A Targeted Analysis of Sulfur Metabolism and Glucosinolate Biosynthesis Pre and Postharvest.

Rocket (Eruca sativa) is a source of health-related metabolites called glucosinolates (GSLs) and isothiocyanates (ITCs) but little is known of the genetic and transcriptomic mechanisms responsible for regulating pre and postharvest accumulations. We present the first de novo reference genome assembly and annotation, with ontogenic and postharvest transcriptome data relating to sulfur assimilation, transport, and utilization. Diverse gene expression patterns related to sulfur metabolism, GSL biosynthesis, and glutathione biosynthesis are present between inbred lines of rocket. A clear pattern of differential expression determines GSL abundance and the formation of hydrolysis products. One breeding line sustained GSL accumulation and hydrolysis product formation throughout storage. Multiple copies of MYB28, SLIM1, SDI1, and ESM1 have increased and differential expression postharvest, and are associated with GSLs and hydrolysis product formation. Two glucosinolate transporter gene (GTR2) copies were found to be associated with increased GSL accumulations in leaves. Monosaccharides (which are essential for primary metabolism and GSL biosynthesis, and contribute to the taste of rocket) were also quantified in leaves, with glucose concentrations significantly correlated with the expression of numerous GSL-related genes. Significant negative correlations were observed between the expression of glutathione synthetase (GSH) genes and those involved in GSL metabolism. Breeding line "B" showed increased GSH gene expression and low GSL content compared to two other lines where the opposite was observed. Co-expression analysis revealed senescence (SEN1) and oxidative stress-related (OXS3) genes have higher expression in line B, suggesting that postharvest deterioration is associated with low GSL concentrations.

The Relationship between Glucosinolates and the Sensory Characteristics of Steamed-Pureed Turnip (Brassica Rapa subsp. Rapa L.).

Glucosinolates (GSLs) are phytochemical compounds that can be found in Brassica vegetables. Seven separate batches of steamed-pureed turnip were assessed for GSL content using liquid chromatography mass spectrometry (LC-MS) and for sensory attributes by sensory profiling (carried out by a trained sensory panel). Twelve individual GSLs, which included 7 aliphatic, 4 indole and 1 arylaliphatic GSL, were identified across all batches. There were significant differences in individual GSL content between batches, with gluconasturtiin as the most abundant GSL. The total GSL content ranged from 16.07 to 44.74 µmol g-1 dry weight (DW). Sensory profiling concluded there were positive correlations between GSLs and bitter taste and negative correlations between GSLs (except glucobrassicanapin) and sweet taste. The batches, which had been purchased across different seasons, all led to cooked turnip that contained substantial levels of GSLs which were subsequently all rated as bitter.

Red Light Is Effective in Reducing Nitrate Concentration in Rocket by Increasing Nitrate Reductase Activity, and Contributes to Increased Total Glucosinolates Content.

Rocket cultivation is increasing to supply the expanding ready-to-eat market because of its unique taste, but crops are often over fertilized to avoid nitrogen deficiencies. This leads to nitrate accumulation in leaves, and the products of their degradation (nitrites and nitrosamines) have been related to several health problems. Nitrate concentrations in rocket and other leafy vegetables are subject to limits by the EU legislation, yet rocket holds a great nutritional value. Degradation products of glucosinolates (isothiocyanates) have been consistently linked with benefits to human health. We investigated the influence of nitrogen application (1 and 8 mM), species [Eruca sativa (L.) Cav. and Diplotaxis tenuifolia (L.) DC.] and light spectrum (full spectrum, red, blue and red + blue) on the nitrate concentration, nitrate reductase activity and glucosinolate content of rocket grown in a soil-less system. Red light decreased the nitrate concentration with respect to the blue spectrum (4,270 vs. 7,100 mg⋅kg-1 of fresh weight, respectively), but such reduction was influenced by the species and the nitrogen level (significantly higher in D. tenuifolia and with the higher concentration of N). The nitrate reductase activity increased under red light in D. tenuifolia, with the lower N concentration. Rocket is known to contain several health-promoting compounds mainly antioxidants and glucosinolates, as secondary metabolites that act as part of plant defense mechanisms. The total content of glucosinolates was mainly affected by the species (D. tenuifolia showed the highest concentrations). Our results will help growers to tailor light spectra with the aim of reducing nitrate concentration and to remain within EU legislative limits, without any detrimental influence on other qualitative parameters in rocket.

Growth temperature influences postharvest glucosinolate concentrations and hydrolysis product formation in first and second cuts of rocket salad.

Rocket salad species (Diplotaxis tenuifolia and Eruca sativa; also known as E. vesicaria) are known for their high concentrations of health-related isothiocyanates, which are derived from secondary metabolites called glucosinolates. Increases in temperature due to climate change and extreme weather event frequencies over the coming decades are likely to influence not only the growth of leafy vegetables, but also their nutritional density. It is therefore essential to determine the impacts of these in order to mitigate crop losses and nutritional decline in future. Our data show there is a strong influence of pre-harvest growth temperatures on glucosinolate biosynthesis and formation of glucosinolate hydrolysis products postharvest, and that this is genotype dependent. High growth temperature (40 °C) severely retarded germination, growth, regrowth, and survival of rocket plants. Highest glucosinolate concentrations were observed in first and second cuts at 40 °C, but did not correspond to highest isothiocyanate concentrations (observed at 30 °C, second cut). Hydrolysis product formation is proportionately not as great as glucosinolate increases at 40 °C, possibly due to inhibition of enzyme function(s) at higher temperatures. These data indicate that high growth temperatures increase glucosinolate accumulation, but growth and productivity is significantly reduced. Much greater emphasis is needed for breeding cultivars tolerant to high growth temperatures in order to maximise nutritional benefits imparted by temperature stress.

Morphophysiology of Potato (Solanum tuberosum) in Response to Drought Stress: Paving the Way Forward.

The cultivated potato (Solanum tuberosum L.) is currently the third most important food crop in the world and is becoming increasingly important to the local economies of developing countries. Climate change threatens to drastically reduce potato yields in areas of the world where the growing season is predicted to become hotter and drier. Modern potato is well known as an extremely drought susceptible crop, which has primarily been attributed to its shallow root system. This review addresses this decades old consensus, and highlights other, less well understood, morphophysiological features of potato which likely contribute to drought susceptibility. This review explores the effects of drought on these traits and goes on to discuss phenotypes which may be associated with drought tolerance in potato. Small canopies which increase harvest index and decrease evapotranspiration, open stem-type canopies which increase light penetration, and shallow but densely rooted cultivars, which increase water uptake, have all been associated with drought tolerance in the past, but have largely been ignored. While individual studies on a limited number of cultivars may have examined these phenotypes, they are typically overlooked due to the consensus that root depth is the only significant cause of drought susceptibility in potato. We review this work, particularly with respect to potato morphology, in the context of a changing climate, and highlight the gaps in our understanding of drought tolerance in potato that such work implies.

Rocket science: A review of phytochemical & health-related research in Eruca & Diplotaxis species.

Rocket species (Eruca spp. and Diplotaxis spp.) are becoming increasingly important leafy salad crops across the world. Numerous scientific research papers have been published in recent years surrounding the potential health benefits associated with phytochemicals contained in leaves, such as flavonoids and isothiocyanates. Other research of note has also been conducted into the unique taste and flavour properties of leaves, which can be hot, peppery, bitter, and sweet depending upon the genotype and phytochemical composition. While research into these aspects is increasing, some studies lack cohesion and in-depth knowledge of commercial breeding and cultivation practices that makes interpretation and application of results difficult. This review draws together all significant research findings in these crops over the last five years, and highlights areas that require further exploration and/or resolution. We also advise on experimental considerations for these species to allow for more meaningful utilisation of findings in the commercial sector.

Taste and Flavor Perceptions of Glucosinolates, Isothiocyanates, and Related Compounds.

Brassicaceae plants are renowned for their taste, aroma and trigeminal characteristics; predominantly bitter taste, sulfurous aroma, and pungency. Compounds responsible for these sensations include the glucosinolates (GSLs) and their hydrolysis products, particularly isothiocyanates (ITCs), but also sulfur-containing volatile compounds. This article reviews the relative importance of taste and flavor perceptions resulting from such compounds; collating evidence from papers where findings are based on sensory analytical correlations, and those that have extracted specific compounds prior to sensory evaluation. Where specific GSLs impart bitterness and many ITCs impart pungency, this is clearly not true for all GSLs and ITCs. Designing crop improvement strategies for sensory traits based on total GSL content would be flawed, as it does not consider the relative differences in sensory characteristics of different GSLs and ITCs, nor the contribution from other GSL hydrolysis products. In addition, some Brassicaceae plants are consumed raw, whilst others are cooked; this affects not only the hydrolysis of GSLs, but also the generation and release of sulfides. Therefore, in breeding new plant varieties, it is prudent to consider the individual GSLs, the typical cooking conditions the plant is subjected to, enzyme stability, and resultant composition of both GSL hydrolysis products (including ITCs) and sulfides.