Antioxidative Response and Phenolic Content of Young Industrial Hemp Leaves at Different Light and Mycorrhiza.

Due to the increasing presence of industrial hemp (Cannabis sativa L.) and its multiple possibilities of use, the influence of different light and several biopreparations based on beneficial fungi and bacteria on hemp's morphological and physiological properties were examined. Different biopreparations and their combinations were inoculated on hemp seed and/or substrate and grown under blue and white light. A completely randomized block design was conducted in four replications within 30 days. For biopreparation treatment, vesicular arbuscular mycorrhiza (VAM) in combination with Azotobacter chroococum and Trichoderma spp. were inoculated only on seed or both on seed and in the substrate. Generally, the highest morphological parameters (stem, root and plant length) were recorded on plants in white light and on treatment with applied Trichoderma spp., both on seed and substrate. Blue light negatively affected biopreparation treatments, resulting in lower values of all morphological parameters compared to control. Leaves pigments were higher under blue light, as compared to the white light. At the same time, 1-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), flavonoids, total flavanol content and phenolic acids were not influenced by light type. Biopreparation treatments did not significantly influence the leaves' pigments content (Chl a, Chl b and Car), nor the phenolic and flavanol content.

Effect of Wheatgrass Juice on Nutritional Quality of Apple, Carrot, Beet, Orange and Lemon Juice.

Fresh fruit and vegetable juices are commonly consumed as a valuable source of nutrients, while wheatgrass juice is, due to its nutritional value, used as a natural dietary supplement. The main aim of this research was to evaluate the effect of wheatgrass juice addition to apple, beet, carrot, orange, and lemon juice on total and in vitro bioaccessible concentrations of K, Ca, Mg, Mn, Fe, and Zn, vitamin C concentration, total phenolic and flavonoid content, and antioxidant activity. In comparison to other juices, wheatgrass juice had the highest total and in vitro bioaccessible concentrations of Ca, Mg, Mn, Fe, and Zn, while beet juice had the highest K concentration. Lemon and orange juices had the highest vitamin C concentration, while the highest total phenolic and flavonoid content were found in wheatgrass juice. After the addition of wheatgrass juice, Ca, Mg, Mn, and Zn concentration increased in all examined juices, vitamin C concentration increased in apple, beet, and carrot juice, total phenolic content increased in carrot juice, while total flavonoid content increased in apple, carrot, and orange juice. In comparison to the examined juices, wheatgrass juice has better nutritional value, and it could be used in a mixture with other juices to improve their nutritional value.

Selenium Biofortification of Soybean Seeds Influences Physiological Responses of Seedlings to Osmotic Stress.

Climate change poses a serious threat to agricultural production. Water deficit in agricultural soils is one of the consequences of climate change that has a negative impact on crop growth and yield. Selenium (Se) is known to be involved in plant defense against biotic and abiotic stress through metabolic, structural, and physiological activity in higher plants. The aim of this study was to investigate the physiological response of Se-biofortified soybean (Glycine max (L.) Merrill) seedlings under osmotic stress. For this research, we used biofortified soybean grain obtained after foliar Se biofortification in 2020. The experiment was conducted in a growth chamber with two cultivars (Lucija and Sonja) grown on filter paper in three replicates. The experiment was carried out with two watering treatments: distilled water (PEG-0) and 2.5% polyethylene glycol 6000 (PEG-2.5) on Se-biofortified seeds (Se) and nonbiofortified seeds (wSe). Contents of lipid peroxidation product (LP), free proline (PRO), total phenolic content (TP), ferric reducing antioxidant power (FRAP), and ascorbic acid (AA) were analyzed in 7-days-old seedlings. Significant differences were detected in the Se content of soybean grains between the two cultivars. A milder reaction to PEG-2.5 was observed in cultivar Lucija in both Se and wSe treatments, which might represent the mitigating effects of Se on osmotic stress in this cultivar. Contrarily, in cultivar Sonja, Se adversely affected all analyzed traits in the PEG-2.5 treatment. Ultimately, Se is a pro-oxidant in Sonja, whereas it represents an anti-oxidant in Lucija. In conclusion, different soybean cultivars show contrasting physiological reactions to both osmotic stress and Se. However, the activation of antioxidant pathways in Sonja can also be interpreted as added value in soybean seedlings as a functional food.

Coumarin Derivatives Act as Novel Inhibitors of Human Dipeptidyl Peptidase III: Combined In Vitro and In Silico Study.

Dipeptidyl peptidase III (DPP III), a zinc-dependent exopeptidase, is a member of the metalloproteinase family M49 with distribution detected in almost all forms of life. Although the physiological role of human DPP III (hDPP III) is not yet fully elucidated, its involvement in pathophysiological processes such as mammalian pain modulation, blood pressure regulation, and cancer processes, underscores the need to find new hDPP III inhibitors. In this research, five series of structurally different coumarin derivatives were studied to provide a relationship between their inhibitory profile toward hDPP III combining an in vitro assay with an in silico molecular modeling study. The experimental results showed that 26 of the 40 tested compounds exhibited hDPP III inhibitory activity at a concentration of 10 µM. Compound 12 (3-benzoyl-7-hydroxy-2H-chromen-2-one) proved to be the most potent inhibitor with IC50 value of 1.10 µM. QSAR modeling indicates that the presence of larger substituents with double and triple bonds and aromatic hydroxyl groups on coumarin derivatives increases their inhibitory activity. Docking predicts that 12 binds to the region of inter-domain cleft of hDPP III while binding mode analysis obtained by MD simulations revealed the importance of 7-OH group on the coumarin core as well as enzyme residues Ile315, Ser317, Glu329, Phe381, Pro387, and Ile390 for the mechanism of the binding pattern and compound 12 stabilization. The present investigation, for the first time, provides an insight into the inhibitory effect of coumarin derivatives on this human metalloproteinase.

Validation of flavonoids as potential dipeptidyl peptidase III inhibitors: Experimental and computational approach.

Fifteen flavonoids were studied for their inhibitory activity against human dipeptidyl peptidase III (hDPP III) combining an in vitro assay with an in silico molecular modeling study. All analyzed flavonoids showed inhibitory effects against hDPP III with the IC50 values ranging from 22.0 to 437.2 µm. Our 3D QSAR studies indicate that the presence of hydrophilic regions at a flavonoid molecule increases its inhibitory activity, while the higher percentage of hydrophobic surfaces has negative impact on enzyme inhibition. Furthermore, molecular dynamics (MD) simulations of the complex of hDPP III with one of the most potent inhibitors, luteolin, were performed, and binding mode analysis revealed that the 3' and 4' hydroxyl group on B-ring as well as 5 and 7 hydroxyl group on A-ring helps luteolin to interact with the Asn391, Asn406, Tyr417, His450, Glu451, Val447, Glu512, Asn545, Gln566, and Arg572 residues. The MD results clearly provide valuable information explaining the importance of flavonoid hydroxyl groups in the mechanism for the binding pattern at the active site of hDPP III.

Hydrogen sulfide: environmental factor or signalling molecule?

Hydrogen sulfide (H2S) has traditionally been thought of as a phytotoxin, having deleterious effects on the plant growth and survival. It is now recognized that plants have enzymes which generate H2S, cysteine desulfhydrase, and remove it, O-acetylserine lyase. Therefore, it has been suggested that H2S is considered as a signalling molecule, alongside small reactive compounds such as hydrogen peroxide (H2O2) and nitric oxide (NO). Exposure of plants to low of H2S, for example from H2S donors, is revealing that many physiological effects are seen. H2S seems to have effects on stomatal apertures. Intracellular effects include increases in glutathione levels, alterations of enzyme activities and influences on NO and H2O2 metabolism. Work in animals has shown that H2S may have direct effects on thiol modifications of cysteine groups, work that will no doubt inform future studies in plants. It appears therefore, that instead of thinking of H2S as a phytotoxin, it needs to be considered as a signalling molecule that interacts with reactive oxygen species and NO metabolism, as well as having direct effects on the activity of proteins. The future may see H2S being used to modulate plant physiology in the field or to protect crops from postharvest spoilage.

Hydrogen sulfide effects on stomatal apertures.

Hydrogen sulfide (H(2)S) has recently been reported to be a signaling molecule in plants. It has been well established that is has such roles in animals and it has been suggested that it is included into the group of gasotransmitters. We have recently shown that hydrogen sulfide causes stomatal opening in the model plant Arabidopsis thaliana. H(2)S can be supplied to the plant tissues from donors such as sodium hydrosulfide (NaSH) or more recently from slow release H(2)S donor molecules such as GYY4137. Both give similar effects, that is, they cause stomatal opening. Furthermore both H(2)S donors reduced the accumulation of nitric oxide (NO) induced by abscisic acid (ABA) treatment of leaf tissues. Here similar work has been repeated in a crop plant, Capsium anuum, and similar data has been obtained, suggesting that such effects of hydrogen sulfide on plants is not confined to model species.

The relationship of proline content and metabolism on the productivity of maize plants.

The free proline content in maize ear-leaves, silk and pollen were analyzed in field grown plants which had matured to the pollination stage. Using maize hybrids PR34F02, PR35P12 and PR36B08 field trials were set up at two locations in eastern Croatia in two different years. Two enzymes of proline metabolism were analyzed in the same leaf samples and specific activities of synthetase (P5CS) and proline dehydrogenase (PDH). Plant productivity was evaluated at harvest by the estimation of total and fully developed grain number per ear and per plant, the mean single grain mass, and the mass of grain per plant. The year in which the plants were grown had a very significant effect on the free proline content in the leaf and pollen, as well as on the enzyme activities assayed. The differences between the plants from the two localities were very significant in all tested parameters of plant grain productivity. There was a significant genotype effect on proline content and P5CS total activity in leaf and on all the productivity parameters. Some of the correlations established suggest that the rate of proline synthesis and degradation in maize ear-leaf at pollination might contribute to the final grain production of the maize plant. Multiple regression analyses was used to further analyze the relationship between proline and grain productivity, but it is clear that future work should include other environmental conditions, plant species and organs such as roots.

Assessment of the influence of different sample processing and cold storage duration on plant free proline content analyses.

INTRODUCTION: A method which is widely accepted for the analysis of free proline content in plant tissues is based on the use of 3% sulfosalicylic acid as an extractant, followed by spectrophotometric quantification of a proline-ninhydrin complex in toluene. However, sample preparation and storage may influence the proline actually measured. This may give misleading or difficult to compare data. OBJECTIVE AND METHODOLOGY: To evaluate free proline levels fresh and frozen strawberry (Fragaria × ananassa Duch.) leaves and soybean [Glycine max (L.) Merr.] hypocotyl tissues were used. These were ground with or without liquid nitrogen and proline extracted with sulfosalicylic acid. A particular focus was the influence of plant sample cold storage duration (1, 4 and 12 weeks at -20°C) on tissue proline levels measured. RESULTS: The free proline content analyses, carried out in leaves of Fragaria × ananassa Duch. as well as in hypocotyls of Glycine max (L.) Merr., showed a significant influence of the sample preparation method and cold storage period. Long-term storage of up to 12 weeks at -20°C led to a significant increase in the measured proline in all samples analysed. CONCLUSION: The observed changes in proline content in plant tissue samples stored at -20°C indicate the likelihood of the over-estimation of the proline content if the proline analyses are delayed. Plant sample processing and cold storage duration seem to have an important influence on results of proline analyses. Therefore it is recommended that samples should be ground fresh and analysed immediately.