Melatonin production by rhizobacteria native strains: Towards sustainable plant growth promotion strategies.

In the current context of climate change and water deficit, the selection of native beneficial microorganisms, such as plant growth-promoting rhizobacteria (PGPR), has become a trend for sustainable agriculture due to their ability to improve plant-bacteria interaction with a minimal adverse effect on the soil microbiota compared to commercial PGPR. Until now, the production of phytohormones like melatonin (MT) by native PGPR and their effect on endogenous MT levels in plants have been poorly studied. MT is a ubiquitous phytohormone that protects plants against biotic and abiotic stress by improving the tolerance of stressed plants. In this work, the production of MT by two native PGPR, Enterobacter 64S1 and Pseudomonas 42P4, was evaluated and both PGPR were applied in Arabidopsis thaliana plants grown under drought conditions to assess the inoculation effects. Parameters such as plant growth, leaf cellular membrane damage, leaf protective compounds, and endogenous MT levels under drought and irrigation conditions were evaluated. The results demonstrated that the native strains Pseudomonas 42P4 and Enterobacter 64S1 produce MT and increase the content of endogenous MT in A. thaliana plants under drought. These native strains improved the tolerance of arabidopsis plants to drought by preventing oxidative and membrane damages and improving plant growth. To the best of our knowledge, this is the first report on MT production by native PGPR and their effects on endogenous MT levels in arabidopsis plants, setting the bases to elucidate the role of native PGPR on water deficit conditions.

Enhanced electrochemical detection of quercetin by Natural Deep Eutectic Solvents.

New trends in analytical chemistry encourage the development of smart techniques and methods aligned with Green Chemistry. In this sense, Natural Deep Eutectic Solvents represents an excellent opportunity as a new generation of green solvents. In this work a new application for them has been proposed and demonstrated. These solvents were synthesized by combinations of inexpensive and natural components like, Glucose, Fructose, Citric acid and Lactic acid. The different natural solvents were easily prepared and added to buffer solution in different concentrations, allowing the enhancement of electrochemical detection of an important representative antioxidant like quercetin (QR) with improved signal up to 380%. QR is a ubiquitous flavonoid widespread in plants and food of plant origin. The proposed method using phosphate buffer with a eutectic mixture of Citric acid, Glucose and water in combination with carbon screen printed electrodes exhibited a good analytical performance. Detection and quantification limits were of 7.97 and 26.3 nM respectively; and repeatability with %RSDs of 1.41 and 7.49 for peak potential and intensity respectively. In addition, it has proved to be faster, greener and cheaper than other sensors and chromatographic methods available with the additional advantage of being completely portable. Furthermore, the obtained results demonstrated that the proposed method is able for the determination of QR in complex food samples.

Melatonin in Arabidopsis thaliana acts as plant growth regulator at low concentrations and preserves seed viability at high concentrations.

Since the discovery of melatonin in plants, several roles have been described for different species, organs, and developmental stages. Arabidopsis thaliana, being a model plant species, is adequate to contribute to the elucidation of the role of melatonin in plants. In this work, melatonin was monitored daily by UHPLC-MS/MS in leaves, in order to study its diurnal accumulation as well as the effects of natural and artificial light treatments on its concentration. Furthermore, the effects of exogenous application of melatonin to assess its role in seed viability after heat stress and as a regulator of growth and development of vegetative tissues were evaluated. Our results indicate that melatonin contents in Arabidopsis were higher in plants growing under natural radiation when compared to those growing under artificial conditions, and its levels were not diurnally-regulated. Exogenous melatonin applications prolonged seed viability after heat stress conditions. In addition, melatonin applications retarded leaf senescence. Its effects as growth promoter were dose and tissue-dependent; stimulating root growth at low concentrations and decreasing leaf area at high doses.

Microchip electrophoresis-single wall carbon nanotube press-transferred electrodes for fast and reliable electrochemical sensing of melatonin and its precursors.

In the current work, single-wall carbon nanotube press-transferred electrodes (SW-PTEs) were used for detection of melatonin (MT) and its precursors tryptophan (Trp) and serotonin (5-HT) on microchip electrophoresis (ME). SW-PTEs were simply fabricated by press transferring a filtered dispersion of single-wall carbon nanotubes on a nonconductive PMMA substrate, where single-wall carbon nanotubes act as exclusive transducers. The coupling of ME-SW-PTEs allowed the fast detection of MT, Trp, and 5-HT in less than 150 s with excellent analytical features. It exhibited an impressive antifouling performance with RSD values of ≤2 and ≤4% for migration times and peak heights, respectively (n = 12). In addition, sample analysis was also investigated by analysis of 5-HT, MT, and Trp in commercial samples obtaining excellent quantitative and reproducible recoveries with values of 96.2 ± 1.8%, 101.3 ± 0.2%, and 95.6 ± 1.2% for 5-HT, MT, and Trp, respectively. The current novel application reveals the analytical power of the press-transfer technology where the fast and reliable determination of MT and its precursors were performed directly on the nanoscale carbon nanotube detectors without the help of any other electrochemical transducer.

Analytical tools for elucidating the biological role of melatonin in plants by LC-MS/MS.

Melatonin (MT) presence in higher plants was recently discovered and the knowledge of its function in vivo is limited. Several studies have recently shown the occurrence of MT and related compounds in grapes and wines. The analysis of MT in plants and foods represents a highly challenging task due to its wide concentration range, the difficulty in the selection of the extraction solvents because of its amphipathic nature, and the fact that it reacts quickly with other matrix components. Thus, sample processing factors; preparation/cleanup procedures; and chromatographic/detection parameters, such as HILIC and reverse phase (C(8) and C(18)) chromatographic modes, ESI, and atmospheric pressure chemical ionization (APCI) in both negative and positive modes were evaluated. Taken together, we have demonstrated that optimal conditions were quite different for each of the matrices under study. A sonication-mediated extraction step was necessary for grape skin (100% v/v methanol) and plant tissues (50% v/v methanol), while wine and must required a SPE preconcentration step. HILIC-(+) APCI ionization was better for MT standards, while C(8) -(+) APCI was the best choice for grape skin and C(18) -(+ESI) was suitable for wine. On the other hand, C(8) -(+)ESI was the most appropriate for vegetal tissues of Arabidopsis thaliana. Proposed methods were validated and the LODs were in the low picogram levels range. The optimized approaches were applied to the determination of MT and its isomer in different vegetal/food samples; levels found within the range: 4.9-440 ng/g.

Monitoring melatonin and its isomer in Vitis vinifera cv. Malbec by UHPLC-MS/MS from grape to bottle.

Several studies have shown the presence of melatonin and related compounds in grapes and wines. The latter provides evidence of the possibility to enhance the nutraceutical properties of premium wines. However, there are many external factors that can influence the levels of this indolamine in grape and wines. In this study, the monitoring of melatonin and its tentatively identified isomer was carried out during the entire winemaking process in Vitis vinifera cv. Malbec by ultra high-performance liquid chromatography-tandem mass spectrometry. Laboratory and pilot studies were carried out to elucidate the role of grape, yeasts, and tryptophan in the evolution of the indolamines during the fermentation process. Melatonin was detected in grape extract within the range 120-160 ng/g while its isomer was found in musts and finished wines. Our results demonstrate that Saccaromyces cervisiae plays a decisive role in contributing to the content of melatonin and its isomer in wine.