Modulation of cherry tomato performances in response to molybdenum biofortification and arbuscular mycorrhizal fungi in a soilless system.

Molybdenum (Mo) is a crucial microelement for both, humans and plants. The use of agronomic biofortification techniques can be an alternative method to enhance Mo content in vegetables. Concomitantly, arbuscular mycorrhizal fungi (AMF) application is a valuable strategy to enhance plant performances and overcome plant abiotic distresses such as microelement overdose. The aim of this research was to estimate the direct and/or indirect effects of Mo supply at four doses [0.0, 0.5 (standard dose), 2.0 or 4.0 µmol L-1], alone or combined with AMF inoculation, on plant performances. In particular, plant height and first flower truss emission, productive features (total yield, marketable yield and average marketable fruit weight) and fruit qualitative characteristics (fruit dry matter, soluble solids content, titratable acidity, ascorbic acid, lycopene, polyphenol, nitrogen, copper, iron and molybdenum) of an established cherry tomato genotype cultivated in soilless conditions were investigated. Moreover, proline and malondialdehyde concentrations, as well as Mo hazard quotient (HQ) in response to experimental treatments were determined. A split-plot randomized experimental block design with Mo dosages as plots and +AMF or -AMF as sub-plots was adopted. Data revealed that AMF inoculation enhanced marketable yield (+50.0 %), as well as some qualitative traits, such as fruit soluble solids content (SSC) (+9.9 %), ascorbic acid (+7.3 %), polyphenols (+2.3 %), and lycopene (+2.5 %). Molybdenum application significantly increased SSC, polyphenols, fruit Mo concentration (+29.0 % and +100.0 % in plants biofortified with 2.0 and 4.0 µmol Mo L-1 compared to those fertigated with the standard dose, respectively) and proline, whereas it decreased N (-25.0 % and -41.6 % in plants biofortified with 2.0 and 4.0 µmol Mo L-1 compared to those fertigated with the standard dose, respectively). Interestingly, the application of AMF mitigated the detrimental effect of high Mo dosages (2.0 or 4.0 µmol L-1). A pronounced advance in terms of plant height 45 DAT, fruit lycopene concentration and fruit Fe, Cu and Mo concentrations was observed when AMF treatment and Mo dosages (2.0 or 4.0 µmol Mo L-1) were combined. Plants inoculated or not with AMF showed an improvement in the hazard quotient (HQ) in reaction to Mo application. However, the HQ - for a consumption of 200 g day-1 of biofortified cherry tomato - remained within the safety level for human consumption. This study suggests that Mo-implementation (at 2.0 or 4.0 µmol L-1) combined with AMF inoculation could represent a viable cultivation protocol to enhance yield, produce premium quality tomato fruits and, concomitantly, improve Mo dose in human diet. In the light of our findings, further studies on the interaction between AMF and microelements in other vegetable crops are recommended.

Humic acid and grafting as sustainable agronomic practices for increased growth and secondary metabolism in cucumber subjected to salt stress.

Salinity stress poses a significant treat to crop yields and product quality worldwide. Application of a humic acid bio stimulant and grafting onto tolerant rootstocks can both be considered sustainable agronomic practices that can effectively ameliorate the negative effects of salinity stress. This study aimed to assess the above mentioned ameliorative effects of both practices on cucumber plants subjected to saline environments. To attain this goal a factorial experiment was carried out in the form of a completely randomized design with three replications. The three factors considered were (a) three different salinity levels (0, 5, and 10 dS m-1 of NaCl), (b) foliar application of humic acid at three levels (0, 100, and 200 mg L-1), and (c) both grafted and ungrafted plants. Vegetative traits including plant height, fresh and dry weight and number of leaf exhibited a significant decrease under increasing salinity stress. However, the application of humic acid at both levels mitigated these effects compared to control plants. The reduction in relative water content (RWC) of the leaf caused by salinity, was compensated by the application of humic acid and grafting. Thus, the highest RWC (86.65%) was observed in grafting plants with 0 dS m-1 of NaCl and 20 mg L-1 of humic acid. Electrolyte leakage (EL) increased under salinity stress, but the application of humic acid and grafting improved this trait and the lowest amount of EL (26.95%) was in grafting plants with 0 dS m-1 of NaCl and 20 mg L-1 of humic acid. The highest amount of catalase (0.53 mmol H2O2 g-1 fw min-1) and peroxidase (12.290 mmol H2O2 g-1 fw min-1) enzymes were observed in the treatment of 10 dS m-1 of NaCl and 200 mg L-1 humic acid. The highest amount of total phenol (1.99 mg g-1 FW), total flavonoid (0.486 mg g-1 FW), total soluble carbohydrate (30.80 mg g-1 FW), soluble protein (34.56 mg g-1 FW), proline (3.86 µg g-1 FW) was in grafting plants with 0 dS m-1 of NaCl and 200 mg L-1 of humic acid. Phenolic acids and phenylalanine ammonia lyase (PAL) and polyphenol oxidase (PPO) enzymes increased with increasing salinity and humic acid levels. Contrary to humic acid, salt stress increased the sodium (Na+) and chlorine (Cl-) and decreased the amount of potassium (K+) and calcium (Ca2+) in the root and leaf of ungrafted cucumber. However, the application 200 mg L-1 humic acid appeared to mitigate these effects, thereby suggesting a potential role in moderating physiological processes and improving growth of cucumber plants subjected to salinity stress. According to the obtained results, spraying of humic acid (200 mg L-1) and the use of salt resistant rootstocks are recommended to increase tolerance to salt stress in cucumber. These results, for the first time, clearly demonstrated that fig leaf gourd a new highly salt-tolerant rootstock, enhances salt tolerance and improves yield and quality of grafted cucumber plants by reducing sodium transport to the shoot and increasing the amount of compatible osmolytes.

Exogenous application of melatonin protects bean and tobacco plants against ozone damage by improving antioxidant enzyme activities, enhancing photosynthetic performance, and preventing membrane damage.

Ozone (O3) pollution is harmful to plants and ecosystems. Several chemicals have been evaluated to protect plants against O3 deleterious effects. However, they are not adequately efficient and/or the environmental safety of their application is questioned. Hence, new chemicals that provide sufficient protection while being safer for environmental application are needed. This study investigates the response of two O3-sensitive plant species (Phaseolus vulgaris L. cv. Pinto and Nicotiana tabacum L. cv. Bel-W3) leaf-sprayed with deionized water (W, control), ethylenediurea (EDU, 1 mM) or melatonin at lower (1 mM) or higher (3 mM) concentrations (Mel_L and Mel_H, respectively), and then exposed to a square wave of 200 ppb O3, lasting 1 day (5 h day-1) for bean and 2 days (8 h day-1) for tobacco. In both species, the photosynthetic activity of O3-exposed plants was about halved. O3-induced membrane damage was also confirmed by increased malondialdehyde (MDA) byproducts compared to control (W). In EDU- and Mel-treated bean plants, the photosynthetic performance was not influenced by O3, leading to reduction of the incidence and severity of O3 visible injury. In bean plants, Mel_L mitigated the detrimental effect of O3 by boosting antioxidant enzyme activities or osmoprotectants (e.g. abscisic acid, proline, and glutathione transferase). In Mel_L-sprayed tobacco plants, O3 negatively influenced the photosynthetic activity. Conversely, Mel_H ameliorated the O3-induced oxidative stress by preserving the photosynthetic performance, preventing membrane damage, and reducing the visible injuries extent. Although EDU performed better, melatonin protected plants against O3 phytotoxicity, suggesting its potential application as a bio-safer and eco-friendlier phytoprotectant against O3. It is worth noting that the content of melatonin in EDU-treated plants remained unchanged, indicating that the protectant mode of action of EDU is not Mel-related.

Impact of Ecklonia maxima Seaweed Extract and Mo Foliar Treatments on Biofortification, Spinach Yield, Quality and NUE.

Seaweed extract (SE) application is a contemporary and sustainable agricultural practice used to improve yield and quality of vegetable crops. Plant biofortification with trace element is recognized as a major tool to prevent mineral malnourishment in humans. Mo deficiency causes numerous dysfunctions, mostly connected to central nervous system and esophageal cancer. The current research was accomplished to appraise the combined effect of Ecklonia maxima brown seaweed extract (SE) and Mo dose (0, 0.5, 2, 4 or 8 µmol L-1) on yield, biometric traits, minerals, nutritional and functional parameters, as well as nitrogen indices of spinach plants grown in a protected environment (tunnel). Head fresh weight (FW), ascorbic acid, polyphenols, N, P, K, Mg and nitrogen use efficiency (NUE) were positively associated with SE treatment. Moreover, head FW, head height (H), stem diameter (SD), ascorbic acid, polyphenols, carotenoids as well as NUE indices were enhanced by Mo-biofortification. A noticeable improvement in number of leaves (N. leaves), head dry matter (DM) and Mo concentration in leaf tissues was observed when SE application was combined with a Mo dosage of 4 or 8 µmol L-1. Overall, our study highlighted that E. maxima SE treatment and Mo supply can improve both spinach production and quality via the key enzyme activity involved in the phytochemical homeostasis of SE and the plant nutritional status modification resulting in an enhanced spinach Mo tolerance.

Effects of ozone and ammonium sulfate on cauliflower: Emphasis on the interaction between plants and insect herbivores.

Ammonium sulfate [(NH4)2SO4] deposition and elevated ozone (O3) concentrations may negatively affect plants and trophic interactions. This study aimed to evaluate for the first time the interactive effects of high (NH4)2SO4 load and elevated O3 levels on cauliflower (Brassica oleracea L.) under field conditions. Cauliflower seedlings were treated with 0 (AS0) or 50 (AS50) kg ha-1 (NH4)2SO4 and exposed to ambient (AOZ, ≈20 ppb) or elevated (EOZ, ≈55 ppb) O3 for about one month, in a Free Air O3 Concentration Enrichment (FACE) system. The oligophagous diamondback moth (Plutella xylostella Linnaeus, 1758) showed a clear preference towards the seedlings treated with AS50, which intensively grazed. Plant-herbivore interactions were driven by (NH4)2SO4 availability, rather than O3, via increased nitrogen content in the leaves. Further laboratory bioassays were followed to confirm the validity of these observations using polyphagous Eri silkmoth larvae (Samia ricini) as a biological model in a standardized experimental setup. Choice assays, where larvae could select leaves among leaf samples from the different experimental conditions, and no-choice assays, where larvae could graze leaves from just one experimental condition, were conducted. In the choice assay, the larvae preferred AS50-treated leaves, in agreement with the field observations with diamondback moth. In the no-choice assay, larval body mass growth was inhibited when fed with leaves treated with EOZ and/or AS50. Larvae fed with AS50-treated leaves displayed increased mortality. These observations coincide with higher NO3 and Zn content in AS50-treated leaves. This study shows that plant-herbivore interactions can be driven by (NH4)2SO4 availability, independently of O3, and suggests that high N deposition may have severe health implications in animals consuming such plant tissues. Key message: Plant-herbivore interactions are driven by high (NH4)2SO4 availability, independently of O3.

Chemical composition and antioxidant activity of Cichorium spinosum L. leaves in relation to developmental stage.

In the present study, chemical composition and bioactivity of Cichorium spinosum leaves were examined at different growth stages for optimum harvest stage and end-use assessment. Total fresh weight and number of leaves were higher at 4th growth stage; however, at this stage the end-use is not indicated for raw consumption but mostly for pickled or dried products. Regarding chemical composition, the highest content of tocopherols and minerals was observed in the 1st growth stage, whereas sugars content was the highest in 3rd and 4th growth stage. Polyunsaturated fatty acids content was the highest during the first two growth stages, whereas antioxidant activity, phenolic acids and total phenolic compounds content in the 3rd stage. In conclusion, the results of this study indicate that chemical composition of C. spinosum is highly depended on development stage, and harvest stage should be considered for alternative uses of end-products with high bioactivity.

Successive harvesting affects yield, chemical composition and antioxidant activity of Cichorium spinosum L.

In the present study, the effect of successive harvesting on yield, chemical composition and antioxidant activity of Cichorium spinosum plants was examined. C. spinosum plants were grown from seeds sown in seed trays containing peat and young seedlings were transplanted in 2L pots containing peat and perlite (1:1v/v). Plants were harvested two or three times during two consecutive growing periods. Total fresh weight and number of leaves were higher for successive harvests in both growing periods comparing to a single harvest. The application of more than two harvests resulted in quality loss during the 1st growing period, while in the 2nd growing period the overall chemical composition, antioxidant properties and phenolic compounds content was higher than the 1st period. In conclusion, cultivation practices such as sowing date and successive harvesting may be useful tools towards the production of high quality end-product with increased bioactive properties without compromising total yield.

A novel symbiovar (aegeanense) of the genus Ensifer nodulates Vigna unguiculata.

BACKGROUND: Cowpea (Vigna unguiculata) forms nitrogen-fixing root nodules with diverse symbiotic bacteria, mainly slow-growing rhizobial species belonging to the genus Bradyrhizobium, although a few studies have reported the isolation of fast-growing rhizobia under laboratory and field conditions. Although much research has been done on cowpea-nodulating bacteria in various countries around the world, very limited information is available on cowpea rhizobia in European soils. The aim of this study was to study the genetic and phenotypic diversity of indigenous cowpea-nodulating rhizobia in Greece. RESULTS: The genetic diversity of indigenous rhizobia associated with cowpea was investigated through a polyphasic approach. ERIC-PCR based fingerprinting analysis grouped the isolates into three groups. Based on the analysis of the 16S rRNA genes, IGS and on the concatenation of six housekeeping genes (recA, glnII, gyrB, truA, thrA and SMc00019), rhizobial isolates were classified within the species Ensifer fredii. However, symbiotic gene phylogenies, based on nodC, nifH and rhcRST genes, showed that the Ensifer isolates are markedly diverged from type and reference strains of E. fredii and formed one clearly separate cluster. The E. fredii strains were able to nodulate and fix nitrogen in cowpea but not in soybean and common bean. CONCLUSION: The present study showed that cowpea is nodulated under field conditions by fast-growing rhizobia belonging to the species E. fredii. Based on the phylogenies, similarity levels of symbiotic genes and the host range, the Ensifer isolates may constitute a new symbiovar for which the name 'aegeanense' is proposed. © 2017 Society of Chemical Industry.

Phylogenetic multilocus sequence analysis of indigenous slow-growing rhizobia nodulating cowpea (Vigna unguiculata L.) in Greece.

Cowpea (Vigna unguiculata) is a promiscuous grain legume, capable of establishing efficient symbiosis with diverse symbiotic bacteria, mainly slow-growing rhizobial species belonging to the genus Bradyrhizobium. Although much research has been done on cowpea-nodulating bacteria in various countries around the world, little is known about the genetic and symbiotic diversity of indigenous cowpea rhizobia in European soils. In the present study, the genetic and symbiotic diversity of indigenous rhizobia isolated from field-grown cowpea nodules in three geographically different Greek regions were studied. Forty-five authenticated strains were subjected to a polyphasic approach. ERIC-PCR based fingerprinting analysis grouped the isolates into seven groups and representative strains of each group were further analyzed. The analysis of the rrs gene showed that the strains belong to different species of the genus Bradyrhizobium. The analysis of the 16S-23S IGS region showed that the strains from each geographic region were characterized by distinct IGS types which may represent novel phylogenetic lineages, closely related to the type species of Bradyrhizobium pachyrhizi, Bradyrhizobium ferriligni and Bradyrhizobium liaoningense. MLSA analysis of three housekeeping genes (recA, glnII, and gyrB) showed the close relatedness of our strains with B. pachyrhizi PAC48T and B. liaoningense USDA 3622T and confirmed that the B. liaoningense-related isolate VUEP21 may constitute a novel species within Bradyrhizobium. Moreover, symbiotic gene phylogenies, based on nodC and nifH genes, showed that the B. pachyrhizi-related isolates belonged to symbiovar vignae, whereas the B. liaoningense-related isolates may represent a novel symbiovar.

Vegetable Organosulfur Compounds and their Health Promoting Effects.

BACKGROUND: Trends in modern pharmaceutical science show an increase in demand for new drugs and diet supplements derived from natural products, while during the last decades, great research is conducted regarding the natural compounds and their medicinal and bioactive properties. Organosulfur compounds are present in many plants and their bioactive properties have been used in folk and traditional medicine throughout the centuries. Not until recently, modern science confirmed and revealed the chemical compounds that are responsible for these properties, the chemistry involved in their biosynthesis and the main mechanisms of action. OBJECTIVE: In the present review, the organosulfur compounds of vegetable origin and their health effects are presented, focusing on the chemical composition of their main compounds, their biosynthesis and the mechanisms involved in their health effects. Sulfur compound sources presented include mainly vegetable species belonging to Allium genus and Brassicaceae family. CONCLUSION: Organosulfur compounds of vegetable sources are very important in human diet, and their regular consumption has a beneficial contribution to health and well-being. Moreover, their medicinal properties and therapeutic effects have been described thousands of years ago, while they constitute an important ingredient in traditional medicines and preparations. However, not until recently, the mechanisms of action of organosulfur compounds have started to unravel, with promising results for further exploitation of these significant bioactive compounds in drug development and novel nutraceutical products. This review reveals the numerous biological activities of organosulfur compounds of vegetable origin, while it also presents the results of recent clinical studies and trials.