Salt-Induced Stress Impacts the Phytochemical Composition and Aromatic Profile of Three Types of Basil in a Genotype-Dependent Mode.

Basil (Ocimum basilicum L.) is among the most widely used aromatic plants of Lamiaceae, often grown in areas where salinity is an adverse factor. Most studies on the effect of salinity on basil focused on the influence of salt stress on productive traits, while few reported on how it affects the phytochemical composition and the aroma profile. Three basil cultivars (Dark Opal, Italiano Classico, and Purple Ruffles) were grown hydroponically for 34 days with two nutrient solutions that differed in NaCl concentration [no NaCl (Control) and 60 mM NaCl]. Yield, secondary metabolite concentration (ß-carotene and lutein), antioxidant activity [1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reduction antioxidant power (FRAP)], and aroma profile based on composition of volatile organic compounds (VOCs) were appraised in response to salinity applications. Salt stress significantly reduced fresh yield in Italiano Classico and Dark Opal by 43.34 and 31.69%, respectively, while no effect was observed in Purple Ruffles. Furthermore, the salt-stress treatment increased ß-carotene and lutein concentrations, DPPH, and FRAP activities, and the total nitrogen content of the latter cultivar. CG-MS analysis revealed significant differences in VOCs composition of the basil cultivars, with Italiano Classico and Dark Opal characterized by the predominance of linalool (average 37.52%), which, however, was negatively affected by salinity. In Purple Ruffles, the predominant VOC compound, estragole (79.50%), was not affected by the deleterious effects of NaCl-induced stress.

Zinc biofortification of hydroponically grown basil: Stress physiological responses and impact on antioxidant secondary metabolites of genotypic variants.

Ocimum basilicum L. is an aromatic plant rich in bioactive metabolites beneficial to human health. The agronomic biofortification of basil with Zn could provide a practical and sustainable solution to address Zn deficiency in humans. Our research appraised the effects of biofortification implemented through nutrient solutions of different Zn concentration (12.5, 25.0, 37.5, and 50 µM) on the yield, physiological indices (net CO2 assimilation rate, transpiration, stomatal conductance, and chlorophyll fluorescence), quality, and Zn concentration of basil cultivars 'Aroma 2' and 'Eleonora' grown in a floating raft system. The ABTS, DPPH, and FRAP antioxidant activities were determined by UV-VIS spectrophotometry, the concentrations of phenolic acids by mass spectrometry using a Q Extractive Orbitrap LC-MS/MS, and tissue Zn concentration by inductively coupled plasma mass spectrometry. Although increasing the concentration of Zn in the nutrient solution significantly reduced the yield, this reduction was less evident in 'Aroma 2'. However, regardless of cultivar, the use of the maximum dose of Zn (50 µM) increased the concentration of carotenoids, polyphenols, and antioxidant activity on average by 19.76, 14.57, and 33.72%, respectively, compared to the Control. The significant positive correlation between Zn in the nutrient solution and Zn in plant tissues underscores the suitability of basil for soilless biofortification programs.

Differential Response to NaCl Osmotic Stress in Sequentially Harvested Hydroponic Red and Green Basil and the Role of Calcium.

Basil (Ocimum basilicum L.) is a heterogeneous reservoir of bioactive compounds that provide recognized benefits to human health, rendering it a model aromatic herb. Notwithstanding the application of nutritional stress, such as sodium chloride (NaCl) salinity, which mainly affects the primary metabolism, it also triggers adaptive mechanisms that involve the production of bioactive secondary metabolites. Genotype selection and the exogenous application of calcium chloride (CaCl2) help minimize salinity's suppressive effects on growth. In the present study, we hypothesize that the ratio of different salt types may induce differential responses in the function of preharvest factors in hydroponic basil culture. In this perspective, the stock nutrient solution (Control) was supplemented with 12.5 mm NaCl + 8.33 mm CaCl2 (Moderate Mix), 25 mm NaCl (Moderate NaCl), 25 mm NaCl + 16.66 of CaCl2 (High Mix), or 50 mM of NaCl (High NaCl) with the objective of evaluating the different impact of salinity on yield, sensory quality (color and aroma profile), and the accumulation of minerals and bioactive compounds in two successive harvests of green and red basil cultivars. Although more productive (+39.0% fresh weight) than the red one, the green cultivar exhibited higher susceptibility to salinity, especially under the High Mix and High NaCl treatments. The addition of CaCl2 to the High Mix solution reduced the sodium by 70.4% and increased the total polyphenols by 21.5% compared to the equivalent isomolar solution (High NaCl). The crop performance in terms of fresh and dry yield improved for both cultivars at the second cut. Regardless of cultivar and salt treatment, successive harvests also increased the concentration of phenols and vitamin C (29.7 and 61.5%, respectively) while reducing (-6.9%) eucalyptol, the most abundant aromatic compound in both cultivars. Salinity, as well as the mechanical stress induced by cutting, improved the functional quality of basil. However, the productive responses to the conditions imposed in our work once again highlighted the importance of genetic background. Specifically, CaCl2 in the Moderate Mix solution preserved fresh leaf weight in the most stress-sensitive green cultivar.

Preharvest Nutrient Deprivation Reconfigures Nitrate, Mineral, and Phytochemical Content of Microgreens.

While imparting gastronomic novelty and sensory delight, microgreens also constitute rudimentary leafy greens packed with nutrients and phytochemicals. As such, they comprise an upcoming class of functional foods. However, apart from bioactive secondary metabolites, microgreens also accumulate antinutritive agents such as nitrate, especially under conducive protected cultivation conditions. The current work examined nutrient deprivation before harvest (DBH), applied by replacing nutrient solution with osmotic water for six and twelve days, as a strategy for reducing microgreen nitrate levels in different species (lettuce, mustard, and rocket). The three species were sown on a peat-based substrate, cultivated in a controlled climate chamber, and harvested 18 days after sowing, when the first two true leaves emerged. DBH impact on major constituents of the secondary metabolome, mineral content, colorimetric, and yield traits was appraised. Nitrate and mineral content were determined through ion chromatography, phenolic composition through UHPLC-Q-Orbitrap HRMS, and carotenoid composition through HPLC-DAD. Nutrient deprivation was effective in reducing nitrate content; however, effective treatment duration differed between species and decline was more precipitous in nitrate hyperaccumulating species such as rocket. Quercetin and kaempferol glycosides were the flavonol glycosides most abundant in brassicaceous microgreens, whereas lettuce microgreens were steeped in caffeoyl quinic acid. DBH interacted with species as it increased the total phenolic content of lettuce, decreased that of rocket, but did not affect mustard. Further research to link changes in phenolic composition to the sensory and in vivo bioactive profile of microgreens is warranted. Notably, brief (≤6 days) DBH can be applied across species with moderate or no impact on the phenolic, carotenoid, and mineral composition of microgreens. Brief DBH applications also have limited impact on microgreens' yield and colorimetric traits hence on the commercial value of the product. They can therefore be applied for reducing microgreen nitrate levels without significantly impacting key secondary metabolic constituents and their potential bioactive role.

Ontogenetic Variation in the Mineral, Phytochemical and Yield Attributes of Brassicaceous Microgreens.

Microgreens constitute novel gastronomic ingredients that combine visual, kinesthetic and bioactive qualities. The definition of the optimal developmental stage for harvesting microgreens remains fluid. Their superior phytochemical content against mature leaves underpins the current hypothesis of significant changes in compositional profile during the brief interval of ontogeny from the appearance of the first (S1) to the second true leaf (S2). Microgreens of four brassicaceous genotypes (Komatsuna, Mibuna, Mizuna and Pak Choi) grown under controlled conditions and harvested at S1 and S2 were appraised for fresh and dry yield traits. They were further analyzed for macro- and micromineral content using inductively coupled plasma optical emission spectrometry (ICP-OES), carotenoid content using high-performance liquid chromatography with a diode-array detector (HPLC-DAD), volatile organic compounds using solid-phase microextraction followed by gas chromatography-mass spectrometry (SPME-GC/MS), anthocyanins and polyphenols using liquid chromatography-high resolution-tandem mass spectrometry (LC-MS/MS) with Orbitrap technology and for chlorophyll and ascorbate concentrations, well as antioxidant capacity by spectrophotometry. Analysis of compositional profiles revealed genotype as the principal source of variation for all constituents. The response of mineral and phytochemical composition and of antioxidant capacity to the growth stage was limited and largely genotype-dependent. It is, therefore, questionable whether delaying harvest from S1 to S2 would significantly improve the bioactive value of microgreens while the cost-benefit analysis for this decision must be genotype-specific. Finally, the lower-yielding genotypes (Mizuna and Pak Choi) registered higher relative increase in fresh yield between S1 and S2, compared to the faster-growing and higher-yielding genotypes. Although the optimal harvest stage for specific genotypes must be determined considering the increase in yield against reduction in crop turnover, harvesting at S2 seems advisable for the lower-yielding genotypes.

Changes in the primary and secondary metabolome of male green asparagus (Asparagus officinalis L.) as modulated by sequential harvesting.

Rising temperature and solar radiation drive the mobilization and depletion of crown-deposited metabolites harnessed for asparagus spear regeneration during the spring harvest season. We presently examined how successive same-season harvests impact the sensory, nutritive and bioactive composition of select green asparagus genotypes. Soluble carbohydrates were analyzed by HPLC-RI, organic acids and polyphenols by HPLC-DAD and metals by ion chromatography. Higher sugars and lower acids accentuated sweetness and lower polyphenols contributed to reduced astringency at the beginning of the harvest season. This trend was reversed as the season advanced and spear sensory quality was compromised by declining sugars and rising acids; however, functional quality improved as antioxidant capacity increased along with the concentrations of quercetin-3-O-rutinoside (rutin) and ascorbic acid. The compositional changes presently described were uniform across all genotypes examined and thus contribute toward our understanding of seasonal variation in the sensory and functional quality of this acclaimed health-promoting product.

Genotype and Successive Harvests Interaction Affects Phenolic Acids and Aroma Profile of Genovese Basil for Pesto Sauce Production.

Basil (Ocimum basilicum L.) is an essential ingredient of the Mediterranean cuisine due to its distinctive aroma. Genovese basil leaves are used to prepare "pesto", a condiment that has always caught the interest of consumers and producers. Usually, basil for industrial processing is harvested more than once to extract a higher yield. However, successive cuts can affect quality traits that play a crucial role in defining the product's final sensory profile. This research was aimed to evaluate the impact of cut on the quantitative and qualitative properties of three Genovese basil cultivars (Aroma 2, Eleonora and Italiano Classico) grown in an open field. Nitrate content, phenolic acids and aromatic profile were determined by ion chromatography (IC), high-performance liquid chromatography (HPLC), and gas chromatography coupled to a mass spectrometer (GC/MS) analysis, respectively. The second harvest increased fresh biomass and total phenolic acids content by 172% and 413%, respectively, with Italiano Classico recording the highest values. The combination of second-cut Aroma 2 yielded the lowest nitrate (473.8 mg kg-1 of fresh weight) and Eugenol (2.4%) levels. In the second harvest, Eleonora showed an increase in eugenol and trans-α-bergamotene of 75.3% and 48.2%, respectively; whereas, eucalyptol and ß-cis-ocimene decreased by 34.4% and 51.6%, respectively. Although successive harvests may increase basil yield and quality overall, the cultivar-dependent response to successive cuts needs to be accounted for in order to accomplish standardization of industrial "pesto" sauce.

Configuration of the Volatile Aromatic Profile of Carob Powder Milled From Pods of Genetic Variants Harvested at Progressive Stages of Ripening From High and Low Altitudes.

Carob powder is increasingly valued as a substitute for cocoa and as a flavor-enhancing component of processed foods. However, little is known about the impact of preharvest factors such as fruit maturity, genotype and altitude on its volatile organic compounds (VOCs) composition. The current study examined the VOCs composition of powder milled from pods of two genotypes cultivated at 15 and 510 m altitude and harvested at six progressive stages of maturity, ranging from fully developed immature green (RS1) to late ripe (RS6). Fifty-six VOCs categorized into acids, esters, aldehydes, ketones, alcohols, furans, and alkanes were identified through HS-SPME GC-MS analysis. Maturity was the most influential factor, followed by altitude and least by genotype. Aldehydes and alcohols correlated positively (r = 0.789; p < 0.001), both accumulated in immature carobs and decreased with progressive ripening, resulting in the attenuation of green grassy aroma. Conversely, acids increased with ripening and dominated the carob volatilome at full maturity, correlating negatively with aldehydes and alcohols (r = -0.835 and r = -0.950, respectively; p < 0.001). The most abundant VOC throughout ripening (17.3-57.7%) was isobutyric acid, responsible for the characteristic cheesy-acidic-buttery aroma of carob powder. The pleasurable aroma detected at the immature stages (RS2 and RS3) was traced to isobutyrate and methyl isobutyrate esters, rendering unripe green carob powder a potential admixture component for improving the aroma of novel food products. Lower altitude favored the accumulation of acids linked to less pleasant aroma, whereas isobutyric acid was more abundant at higher altitude. This constitutes a significant indication that higher altitude enhances the characteristic carob-like aroma and sensory quality of carob powder.

Phenolic Constitution, Phytochemical and Macronutrient Content in Three Species of Microgreens as Modulated by Natural Fiber and Synthetic Substrates.

The present study examined the modulatory effects of natural fiber substrates (agave fiber, coconut fiber and peat moss) and synthetic alternatives (capillary mat and cellulose sponge) on the nutritive and phytochemical composition of select microgreens species (coriander, kohlrabi and pak choi) grown in a controlled environment. Polyphenols were analyzed by UHPLC-Q-Orbitrap-HRMS, major carotenoids by HPLC-DAD, and macro-minerals by ion chromatography. Microgreens grown on peat moss had outstanding fresh and dry yield but low dry matter content. Natural fiber substrates increased nitrate and overall macro-mineral concentrations in microgreens compared to synthetic substrates. The concentrations of chlorophylls, carotenoids and ascorbate were influenced primarily by species. On the contrary, variability in polyphenols content was wider between substrates than species. Out of twenty phenolic compounds identified, chlorogenic acid and quercetin-3-O-rutinoside were most abundant. Hydroxycinnamic acids and their derivatives accounted for 49.8% of mean phenolic content across species, flavonol glycosides for 48.4% and flavone glycosides for 1.8%. Peat moss provided optimal physicochemical conditions that enhanced microgreens growth rate and biomass production at the expense of phenolic content. In this respect, the application of controlled stress (eustress) on microgreens growing on peat moss warrants investigation as a means of enhancing phytochemical composition without substantial compromise in crop performance and production turnover. Finally, nitrate deprivation practices should be considered for microgreens grown on natural fiber substrates in order to minimize consumer exposure to nitrate.

Selenium Biofortification Impacts the Nutritive Value, Polyphenolic Content, and Bioactive Constitution of Variable Microgreens Genotypes.

Selenium (Se) is considered essential for human nutrition as it is involved in the metabolic pathway of selenoproteins and relevant biological functions. Microgreens, defined as tender immature greens, constitute an emerging functional food characterized by overall higher levels of phytonutrients than their mature counterparts. The nutraceutical value of microgreens can be further improved through Se biofortification, delivering Se-enriched foods and potentially an enhanced content of bioactive compounds. The current study defined the effect of sodium selenate applications at three concentrations (0, 8, and 16 µM Se) on the bioactive compounds and mineral content of coriander, green basil, purple basil, and tatsoi microgreens grown in soilless cultivation. Analytical emphasis was dedicated to the identification and quantification of polyphenols by UHPLC-Q-Orbitrap-HRMS, major carotenoids by HPLC-DAD, and macro micro-minerals by ICP-OES. Twenty-seven phenolic compounds were quantified, of which the most abundant were: Chlorogenic acid and rutin in coriander, caffeic acid hexoside and kaempferol-3-O(caffeoyl) sophoroside-7-O-glucoside in tatsoi, and cichoric acid and rosmarinic acid in both green and purple basil. In coriander and tatsoi microgreens, the application of 16 µM Se increased the total phenols content by 21% and 95%, respectively; moreover, it improved the yield by 44% and 18%, respectively. At the same Se dose, the bioactive value of coriander and tatsoi was enhanced by a significant increase in rutin (33%) and kaempferol-3-O(feruloyl)sophoroside-7-O-glucoside (157%), respectively, compared to the control. In green and purple basil microgreens, the 8 µM Se application enhanced the lutein concentration by 7% and 19%, respectively. The same application rate also increased the overall macroelements content by 35% and total polyphenols concentration by 32% but only in the green cultivar. The latter actually had a tripled chicoric acid content compared to the untreated control. All microgreen genotypes exhibited an increase in the Se content in response to the biofortification treatments, thereby satisfying the recommended daily allowance for Se (RDA-Se) from 20% to 133%. The optimal Se dose that guarantees the effectiveness of Se biofortification and improves the content of bioactive compounds was 16 µM in coriander and tatsoi, and 8 µM in green and purple basil.

Genotype-Specific Modulatory Effects of Select Spectral Bandwidths on the Nutritive and Phytochemical Composition of Microgreens.

Advanced analytical data on microgreens' response to different light spectra constitutes a valuable resource for designing future crop-specific spectral management systems. The current study defined variation in productivity, nutritive and functional quality (mineral-carotenoid-polyphenolic profiles and antioxidant capacity) of novel microgreens (amaranth, cress, mizuna, purslane) in response to select spectral bandwidths (red, blue, blue-red), and appraised clustering patterns configured by the genotype-light-spectrum nexus. Growth parameters dependent on primary metabolism were most favored by blue-red light's efficiency in activating the photosynthetic apparatus. Nitrate accumulation was higher under monochromatic light owing to the dependency of nitrite reductase on the light-driven activity of PSI, most efficiently promoted by blue-red light. Although mineral composition was mostly genotype-dependent, monochromatic red and blue lights tended to increase K and Na and decrease Ca and Mg concentrations. Lutein, ß-carotene, and lipophilic antioxidant capacity were generally increased by blue-red light putatively due to the coupling of heightened photosynthetic activity to increased demand for protection against oxidative stress; the disparate response however of purslane highlights the importance of genotype specificity in these responses and calls for additional investigation. Analysis of polyphenols by Orbitrap LC-MS/MS revealed substantial genotypic differences. Most abundant phenolics were chlorogenic acid ( x ¯ = 5503 µg g-1 dw), feruloylquinic acid ( x ¯ = 974.1 µg g-1 dw), and caffeoyl feruloyl tartaric acid ( x ¯ = 993 µg g-1 dw). Hydroxycinnamic acids accounted for 79.0% of the mean total phenolic content across species, flavonol glycosides for 20.7%, and flavone glycosides for 0.3%. The general response across species was a decrease in individual polyphenolic constituents, particularly flavonol glycosides, and total polyphenols under blue-red light. The pronounced effectiveness of monochromatic blue light in eliciting synthesis of flavonoids could be linked to their capacity for absorbing shorter wavelengths thereby quenching generated photo-oxidation potential. The light-induced stimulation of the phenylpropanoid pathway by monochromatic blue light through epigenetic mechanisms or redox signaling in the photosynthetic apparatus warrants further investigation. The current work highlights how optimized genetic background combined with effective light management might facilitate the production of superior functional quality microgreens.

The occurrence of nitrate and nitrite in Mediterranean fresh salad vegetables and its modulation by preharvest practices and postharvest conditions.

Winter and summer nitrate/nitrite concentrations in 11 salad vegetables were surveyed using a validated HPLC-DAD method. Nitrate was highest in rocket, both in winter (x̅ = 3974 mg kg-1 fw) and summer (x̅ = 3819 mg kg-1 fw). High nitrate accumulators included spinach, purslane, chards, dill, coriander and parsley. Wide intra-species variability and levels in excess of permitted maxima highlighted the importance of monitoring vegetable production methods to protect consumer health. Occurrence of detectible nitrite (14-352 mg kg-1 fw) was most frequent in winter head cabbage. Three additional experiments examined the seasonal effects of nitrogen (N) fertilization rate, application method, formulation and postharvest storage on nitrate and nitrite levels in lettuce, rocket and spinach. Violation of current nitrate limits is likely when total N exceeds 200 kg ha-1, particularly in rocket and spinach. Postharvest nitrate reduction requires exogenous microbial nitrate reductase activity, which is unlikely to be achieved without visible loss of quality.

Functional quality in novel food sources: Genotypic variation in the nutritive and phytochemical composition of thirteen microgreens species.

Compositional variation was examined across 13 microgreens species/subspecies representing Brassicaceae, Chenopodiaceae, Lamiaceae, Malvaceae and Apiaceae, grown in controlled environment. Macro-mineral concentrations were determined by ion chromatography, chlorophyll and ascorbate concentrations, and hydrophilic/lipophilic antioxidant potentials by spectrophotometry, and major carotenoids by HPLC-DAD. Nitrate hyper-accumulators and wide genotypic differences in Na, K and S concentrations were identified. Antioxidant capacity was highest in brassicaceous microgreens and significant genotypic variation was demonstrated in chlorophyll and carotenoid concentrations. High phenolic content was confirmed in Lamiaceae microgreens, with significant varietal differences, and alternative phenolics-rich microgreens from the Apiaceae were identified. Twenty-eight phenolic compounds were variably detected and quantitated through Orbitrap LC-MS/MS with flavonol glycosides, flavones and flavone glycosides, and hydroxycinnamic acids representing 67.6, 24.8 and 7.6% of the mean total phenolic content across species, respectively. The obtained information is critical for selecting new species/varieties of microgreens that may satisfy demand for both taste and health.

Biochemical and histological contributions to textural changes in watermelon fruit modulated by grafting.

Increased watermelon fruit flesh firmness is systematically incurred with grafting on Cucurbita hybrid rootstocks (heterografting). Possible differences in mesocarp cell wall constitution and histology between heterografted, homeografted (self-grafted) and non-grafted watermelon were examined, as well as their contributions to fruit texture. Firmness correlated positively (r=0.78, p<0.001) with cell density (cellsmm-2) which was higher in heterografts (5.83) than homeografts (4.64) and non-grafted controls (4.69). Mean cell size was smallest in heterografts and correlated negatively (r=-0.75, p<0.001) with firmness. Cell wall material, particularly the water-insoluble pectin fractions associated with firmness, were highest in heterografts. No associations with firmness were found for cell wall neutral sugars and membrane permeability. Higher parenchymatic cell density with higher content of alcohol insoluble residue and more abundant water-insoluble pectin fractions underscore enhanced firmness in heterografts. Possible implication of osmolytes in rootstock-mediated cell pressure regulation warrants further investigation.

Configuration of watermelon fruit quality in response to rootstock-mediated harvest maturity and postharvest storage.

BACKGROUND: The configuration of watermelon fruit quality was analysed in a multi-factorial approach accounting for the effects of grafting, harvest maturity and postharvest storage. Diploid, seeded, hybrid cv. Pegasus, cultivated as scion on interspecific hybrid squash rootstock TZ148 and as non-grafted control, was stored at 25 °C following sequential harvests from the onset of ripening to over-maturity. RESULTS: Delayed rootstock-mediated climax in pulp lycopene and chroma was observed, while both were heightened by postharvest storage when harvest preceded full maturity. Pulp firmness was increased by 46.5% on TZ148, while postharvest decrease in firmness was non-significant. Non-grafted fruits attained their peak in pulp carbohydrate content earlier during ripening. Monosaccharide content declined and sucrose content increased both preharvest and postharvest; overall sugar content declined by 4.3% during storage. Pulp acidity decreased steadily with ripening but was moderately increased by grafting. Citrulline content increased by 12.5% on TZ148; moreover, it climaxed with ripening and declined with storage only in grafted fruit. CONCLUSION: Grafting enhances pulp texture and bioactive composition. Potential suppression of sugar content as a result of grafting is minimized at full commercial maturity. Brief postharvest ambient storage enhances pulp lycopene and chroma, especially in early-picked fruit, notwithstanding the depletion of monosaccharides and citrulline and a limited deterioration of texture. © 2015 Society of Chemical Industry.