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The mechanism-based inactivation of four horseradish peroxidase (HRP-C) enzyme variants has been studied kinetically with either hydrogen peroxide or the xenobiotic m-chloroperoxybenzoic acid (mClO2-BzOH) as sole substrate. The concentration and time dependence of inactivation was investigated for the wild-type plant enzyme (HRP-C), the unglycosylated recombinant enzymes (HRP-C*), and two site-directed mutants with Phe143 replaced by Ala ([F143A]HRP-C*) of Arg38 replaced by Lys ([R38K]HRP-C*). The number of turnovers ( r ) of H2O2 required to completely inactivate the enzymes was found to vary between the different enzymes with HRP-C being most resistant to inactivation (r = 625), HRP-C* and [F143A]HRP-C* being approximately twice as sensitive (r = 335 and 385, respectively) in comparison, and [R38K]HRP-C* being inactivated much more easily (r = 20). In the cases of HRP-C* and [F143A]HRP-C*, compared to HRP-C the differences were due to the absence of glycosylation on the exterior of the proteins, whilst the [R38K]HRP-C* variant exhibited a distinct mechanistic difference. When mClO2BzOH was used as the substrate the differences in sensitivity to inactivation disappeared. The values of r were all around 3 reflecting the strong affinity of mClO2BzOH for the active site. The apparent rate constant for inactivation by H2O2 was approximately ten times lower. The affinity of compound I for H2O2 leading to the formation of a transitory intermediate implicated in the inactivation of peroxidase decreased in the order HRP-C, HRP-C*, [F143A]HRP-C*, [R38K]HRP-C*. [ABSTRACT FROM AUTHOR]

Melatonin (N-acetyl-5-methoxytryptamine) is of particular importance as a chronobiological hormone in mammals, acting as a signal of darkness that provides information to the brain and peripheral organs. It is an endogenous synchronizer for both endocrine (i.e., via neurotransmitter release) and other physiological rhythms. In this work we will try to add to the series of scientific events and discoveries made in plants that, surprisingly, confirm the great similarity of action of melatonin in animals and plants. The most relevant milestones on the 25 years of phytomelatonin studies are presented, from its discovery in 1995 to the discovery of its receptor in plants in 2018, suggesting it should be regarded as a new plant hormone. [ABSTRACT FROM AUTHOR]

Water stress is one of the most significant abiotic stresses that disrupts the osmotic balance of plants and consequently reduces their growth and performance. In recent years, it has been found that serotonin, as a signaling and regulatory molecule, can play important roles in the growth and development of plants and enhance their tolerance to abiotic stresses. Saffron is a plant known for its medicinal and culinary properties. Its distinct flavor, aroma, and vibrant color make it a sought-after ingredient in various cuisines and traditional medicines. The aim of this study is to investigate the possible effect of serotonin growth regulator on some morphophysiological and biochemical characteristics of saffron plant under water stress conditions. Water stress was applied using polyethylene glycol 6000 at a level of 30%, w/v. Serotonin was also applied exogenously at a concentration of 100 µM in both foliar and root applications. The experimental findings demonstrated that water stress had a detrimental impact on various growth and photosynthetic parameters including FW, DW, SH, RWC, photosynthetic pigments content, Pn, Fv/Fm, C and Ci. Under these conditions, H 2 O 2 content and ion leakage increased. The increase in the content of proline and sugars also confirmed that the saffron plant was placed in unfavorable growth conditions. Serotonin application in both foliar and root applications and especially root treatment under stressful conditions improved plant growth by activating enzymatic and non-enzymatic antioxidant systems. Overall, the exogenous application of serotonin increased the resistance of saffron plants to water stress., (© 2024. The Author(s).)

Cascade cropping systems (CCS) utilize leachate from a primary crop to grow secondary crops and enhance the efficient use of water and fertilizers in areas with scarce water resources. A preliminary study investigated the effect of melatonin in a cascade cropping system to potentially improve plant tolerance to abiotic stresses. This study aimed to cultivate Salicornia fruticosa in this cropping system to reduce nutrient discharge and assess the impact of exogenous melatonin on Salicornia growth and quality. The CCS included a primary crop of Salicornia grown in an agro-industrial compost or peat. Leachates from these media were used to cultivate the same plant once again in a floating system under four treatments: compost leachate (T1), peat leachate (T2), 100% nutrient solution (NS) (T3), 50% NS (T4) strength. Four concentrations of exogenous melatonin were applied in foliar spray: 0, 100, 200, and 400 µM. Melatonin application increased yield, with the highest values observed when plants were grown in T1. Water use efficiency was also maximized in T1 and with both 200 and 400 µM melatonin applications. The highest nitrogen use efficiency was achieved in plants grown in peat leachate. The lipid membrane damage was assessed revealing that plants grown in compost leachate exhibited the lowest MDA values regardless of melatonin concentrations. The accumulation of some antinutritional compounds (nitrate, oxalate, and sodium) were the highest in those plants grown in compost leachate. Overall, shoots grown in peat leachate exhibited the best phytochemical profile (total phenol content, total flavonoids, and antioxidant capacity), with peak values in plants treated with 200 µM melatonin. These findings suggest that S. fruticosa can be effectively cultivated using leachate from a previous crop in a floating system and that exogenous melatonin application enhances the yield and nutritional quality of Salicornia shoots., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Giménez, Gallegos-Cedillo, Benaissa, Egea-Gilabert, Signore, Ochoa, Gruda, Arnao and Fernández.)

The important role of melatonin in plant growth and metabolism together with recent advances in the potential use of nanomaterials have opened up interesting applications in agriculture. Various nanovehicles have been explored as melatonin carriers in animals, and it is now important to explore their application in plants. Recent findings have substantiated the use of silicon and chitosan nanoparticles (NPs) in targeting melatonin to plant tissues. Although melatonin is an amphipathic molecule, nanocarriers can accelerate its uptake and transport to various plant organs, thereby relieving stress and improving plant shelf-life in the post-harvest stages. We review the scope and biosafety concerns of various nanomaterials to devise novel methods for melatonin application in crops and post-harvest products., Competing Interests: Declaration of interests The authors declare no conflicts of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The presence of melatonin in plants, called phytomelatonin, has gained great interest in recent years. The determination of phytomelatonin levels in plant extracts for both physiological and plant foodstuff studies requires sophisticated techniques due to the low endogenous levels of this indolic compound with hormonal nature. This chapter presents the most common and advanced techniques in the determination of phytomelatonin, with special emphasis on the techniques of extraction, cleaning, separation, detection, identification, and quantification. Multiple examples and recommendations are presented for a clear overview of the pros and cons of phytomelatonin determinations in plant tissues, seeds, and fruits, mainly., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Melatonin, a tryptophan-derived molecule, is endogenously generated in animal, plant, fungal and prokaryotic cells. Given its antioxidant properties, it is involved in a myriad of signaling functions associated with various aspects of plant growth and development. In higher plants, melatonin (Mel) interacts with plant regulators such as phytohormones, as well as reactive oxygen and nitrogen species including hydrogen peroxide (H2O2), nitric oxide (NO) and hydrogen sulfide (H2S). It shows great potential as a biotechnological tool to alleviate biotic and abiotic stress, to delay senescence and to conserve the sensory and nutritional quality of postharvest horticultural products which are of considerable economic importance worldwide. This review provides a comprehensive overview of the biochemistry of Mel, whose endogenous induction and exogenous application can play an important biotechnological role in enhancing the marketability and hence earnings from postharvest horticultural crops., (© The Author(s) 2021. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Biotic stress causes immense damage to agricultural products worldwide and raises the risk of hunger in many areas. Plants themselves tolerate biotic stresses via several pathways, including pathogen-associated molecular patterns (PAMPs), which trigger immunity and plant resistance (R) proteins. On the other hand, humans use several non-ecofriendly methods to control biotic stresses, such as chemical applications. Compared with chemical control, melatonin is an ecofriendly compound that is an economical alternative strategy which can be used to protect animals and plants from attacks via pathogens. In plants, the bactericidal capacity of melatonin was verified against Mycobacterium tuberculosis , as well as multidrug-resistant Gram-negative and -positive bacteria under in vitro conditions. Regarding plant-bacteria interaction, melatonin has presented effective antibacterial activities against phytobacterial pathogens. In plant-fungi interaction models, melatonin was found to play a key role in plant resistance to Botrytis cinerea , to increase fungicide susceptibility, and to reduce the stress tolerance of Phytophthora infestans . In plant-virus interaction models, melatonin not only efficiently eradicated apple stem grooving virus (ASGV) from apple shoots in vitro (making it useful for the production of virus-free plants) but also reduced tobacco mosaic virus (TMV) viral RNA and virus concentration in infected Nicotiana glutinosa and Solanum lycopersicum seedlings. Indeed, melatonin has unique advantages in plant growth regulation and increasing plant resistance effectiveness against different forms of biotic and abiotic stress. Although considerable work has been done regarding the role of melatonin in plant tolerance to abiotic stresses, its role in biotic stress remains unclear and requires clarification. In our review, we summarize the work that has been accomplished so far; highlight melatonin's function in plant tolerance to pathogens such as bacteria, viruses, and fungi; and determine the direction required for future studies on this topic.

Phytomelatonin (plant melatonin) is chemically related to the amino acid tryptophan and has many diverse properties. Phytomelatonin is an interesting compound due to its outstanding actions at the cellular and physiological level, especially its protective effect in plants exposed to diverse stress situations, while its vegetable origin offers many opportunities because it is a natural compound. We present an overview of its origin, its action in plants in general (particularly in plant species with high levels of phytomelatonin), and its possibilities for use as a nutraceutical with particular attention paid to the beneficial effects that it may have in human health. The differences between synthetic melatonin and phytomelatonin, according to its origin and purity, are presented. Finally, the current market for phytomelatonin and its limits and potentials are discussed., Competing Interests: The authors declare no conflict of interest.

Introduction: Melatonin, an indoleamine well known in vertebrates and structurally related to other important substances such as tryptophan or indole-3-acetic acid, is also present in the plant kingdom although its specific function(s) remain to be established. The emerging field of melatonin studies in plants has progressed very slowly, mainly due to the problems associated with melatonin quantification in plants., Objective: Two commonly used procedures for plant samples are compared. The analytical characteristics of both procedures are quantitatively presented using different solvents and small amounts of fresh biological material, and the respective recovery rates and quantitative limits are presented. Some improvements are suggested., Methodology: Two different sample extraction procedures were compared: a direct-sample extraction (DSE) and a homogenised- sample extraction (HSE). Melatonin was then determined in the respective plant samples by HPLC with fluorescence detection., Results: Using the DSE procedure, more than 94% melatonin was recovered from standard solutions, whereas levels higher than 93% were recovered from the spiked plant samples, with little difference between ethyl acetate and chloroform extractions. In the case of HSE, the recoveries of melatonin were approximately half and never higher than 55%. The ultrasonic treatment proposed in the DSE procedure showed different levels of efficiency (2-20%), depending on the sample., Conclusion: This study has established that, with the direct sample extraction procedure, higher recovery rates are obtained both in standard solutions and in plant samples. The straightforwardness and reproducibility of the extraction procedure is accompanied by the high sensitivity obtained with fluorescence detection.

Melatonin (N-acetyl-5-methoxytryptamine) is a biogenic indoleamine structurally related with other important substances such as tryptophan, serotonin, indole-3-acetic acid (IAA). In mammals, birds, reptiles and fish melatonin is a biological modulator of several timing (circadian) processes such as mood, sleep, sexual behavior, immunological status, etc. Since its discovery in plants in 1995 several physiological roles, including a possible role in flowering, circadian rhythms and photoperiodicity and as growth-regulator have been postulated. Recently, a possible role in rhizogenesis in lupin has also been proposed. Here, these actions of melatonin in plant development are commented on and some other interesting recent data concerning melatonin in plants are also discussed. The need for more investigation into melatonin and plants is presented as an obvious conclusion.

The 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid)/H2O2/horseradish peroxidase (ABTS/H2O2/HRP) decoloration method permits the evaluation of the antioxidant activity of complex food samples. This method is capable of determining both hydrophilic (in buffered media) and lipophilic (in organic media) antioxidant properties in the same sample. Its application to the study of the antioxidant properties of grapes, one white and two black, has permitted us to establish important differences in the antioxidant activity. The black grapes showed higher antioxidant activity than the white variety. The determination of hydrophilic antioxidant activity (HAA) and lipophilic antioxidant activity (LAA) in the three grapes showed that a minor contribution (4-13%) of total antioxidant activity (TAA) was due to LAA in all cases. The experimentally determined HAA could be approximated using the relative contributions of the calculated values of reference compounds (total phenols as gallic acid and anthocyanins as malvidin).

The reactions of the fungal enzymes Arthromyces ramosus peroxidase (ARP) and Phanerochaete chrysosporium lignin peroxidase (LiP) with hydrogen peroxide (H(2)O(2)) have been studied. Both enzymes exhibited catalase activity with hyperbolic H(2)O(2) concentration dependence (K(m) approximately 8-10 mm, k(cat) approximately 1-3 s(-1)). The catalase and peroxidase activities of LiP were inhibited within 10 min and those of ARP in 1 h. The inactivation constants were calculated using two independent methods; LiP, k(i) approximately 19 x 10(-3) s(-1); ARP, k(i) approximately 1.6 x 10(-3) s(-1). Compound III (oxyperoxidase) was detected as the majority species after the addition of H(2)O(2) to LiP or ARP, and its formation was accompanied by loss of enzyme activity. A reaction scheme is presented which rationalizes the turnover and inactivation of LiP and ARP with H(2)O(2). A similar model is applicable to horseradish peroxidase. The scheme links catalase and compound III forming catalytic pathways and inactivation at the level of the [compound I.H(2)O(2)] complex. Inactivation does not occur from compound III. All peroxidases studied to date are sensitive to inactivation by H(2)O(2), and it is suggested that the model will be generally applicable to peroxidases of the plant, fungal, and prokaryotic superfamily.