Melatonin (MT; N-acetyI-5-methoxytryptamine) is an amine hormone involved in abiotic stress resistance. Previous studies have confirmed that melatonin can promote seed germination, mediate physiological regulation mechanisms, and stimulate crop growth under stress. However, the osmotic regulation mechanism by which exogenous melatonin mediates salt tolerance in cotton is still largely unknown. To investigate the effect of salt stress on melatonin concentration in germinating cotton seeds, we analyzed melatonin content over time during seed germination under different treatments. Melatonin content reached its minimum at day 6, while cotton germination rates peaked at day 6, indicating melatonin content and seed germination are correlated. Then we investigated the effects of 10–100 μM melatonin treatments on membrane lipid peroxides and osmotic adjustment substances during cotton seed germination under salt stress. Salt stress led to electrolyte leakage (EL) as well as accumulations of hydrogen peroxide (H₂O₂), malondialdehyde (MDA), organic osmotic substances (i.e., proline, soluble sugars), and inorganic osmotic substances (i.e., Na⁺, Cl^-). Meanwhile, the contents of melatonin, soluble proteins, and K⁺ as well as the K⁺/Na⁺ balance decreased, indicating that salt stress inhibited melatonin synthesis and damaged cellular membranes, seriously affecting seed germination. However, melatonin pretreatment at different concentrations alleviated the adverse effects of salt stress on cotton seeds and reduced EL as well as the contents of H₂O₂, MDA, Na⁺, and Cl^-. The exogenous application of melatonin also promoted melatonin, soluble sugar, soluble proteins, proline, and K⁺/Na⁺ contents under salt stress. These results demonstrate that supplemental melatonin can effectively ameliorate the repression of cotton seed germination by enhancing osmotic regulating substances and adjusting ion homeostasis under salt stress. Thus, melatonin may potentially be used to protect cotton seeds from salt stress, with the 20 μM melatonin treatment most effectively promoting cotton seed germination and improving salt stress tolerance.

Although previous studies have found that melatonin can promote seed germination, the phytohormone regulation mechanism by which exogenous melatonin mediates salt tolerance during cotton seed germination is still largely unknown. The effects of melatonin on germination traits and physiological parameters of GXM9 cotton seeds (Gossypium hirsutum L.) under three salt stress treatments (CK, germination of seeds pretreated with water alone; S, germination of seeds pretreated in 150 mM NaCl under salt stress; SM, germination of seeds pretreated in 20 μM melatonin under 150 mM NaCl solution) in the laboratory was investigated. The results showed that salt stress (150 mM) inhibited cotton seed germination and endogenous melatonin accumulation, and pretreatment with 20 μM exogenous melatonin enhanced the cotton germination rate and hypocotyl length as well as the content of endogenous melatonin during seed germination. This suggests that exogenous melatonin promotes seed germination from a morphological perspective. The contents of starch, α-amylase (EC3.3.1.1), β-galactosidase (EC3.2.1.23), abscisic acid (ABA), and gibberellin (GA) were determined simultaneously. The results showed that the α-amylase and β-galactosidase contents in the cotton seeds decreased by 56.97% and 20.18%, respectively, under salt stress compared with the control, while the starch content increased by 11.53% compared with the control at day 7. The ABA content increased by 25.18% and GA content decreased by 27.99% under salt stress compared with the control at 24 h. When exogenous melatonin was applied to the cotton seeds, the content of α-amylase and β-galactosidase increased by 121.77% and 32.76%, respectively, whereas the starch contents decreased by 13.55% compared with the S treatment at day 7. Similarly, the ABA content increased by 12.20% and the GA content increased by 4.77% at 24 h. To elucidate the molecular mechanism by which melatonin promotes seed germination under salt stress, the effects of ABA- and GA-related genes on plant hormone signal transduction were analyzed by quantitative real-time PCR and RNA sequencing. The results indicated that melatonin regulated the expression of ABA and GA genes in the plant signal transduction pathway, induced embryo root development and seed germination, and alleviated dormancy. The expression of the ABA signaling gene GhABF2 was up-regulated and GhDPBF2 was down-regulated, and the expression of GA signaling genes (e.g., GhGID1C and GhGID1B) was up-regulated by melatonin. In conclusion, melatonin enhances salt tolerance in cotton seeds by regulating ABA and GA and by mediating the expression of hormone-related genes in plant hormone signal transduction. This should help us to explore the regulatory mechanisms of cotton resistance and provide a foundation for the cultivation of new varieties.

Background: As damage to the ecological environment continues to increase amid unreasonable amounts of irrigation, soil salinization has become a major challenge to agricultural development. Melatonin (MT) is a pleiotropic signal molecule and indole hormone, which alleviates the damage of abiotic stress to plants. MT has been confirmed to eliminate reactive oxygen species (ROS) by improving the antioxidant system and reducing oxidative damage under adversity. However, the mechanism by which exogenous MT mediates salt tolerance by regulating the photosynthetic capacity and ion balance of cotton seedlings still remains unknown. In this study, the regulatory effects of MT on the photosynthetic system, osmotic modulators, chloroplast, and anatomical structure of cotton seedlings were determined under 0–500 μM MT treatments with salt stress induced by treatment with 150 mM NaCl. Results: Salt stress reduces the chlorophyll content, net photosynthetic rate, stomatal conductance, intercellular CO₂ concentration, transpiration rate, PSII photochemical efficiency, PSII actual photochemical quantum yield, the apparent electron transfer efficiency, stomata opening, and biomass. In addition, it increases non-photochemical quenching. All of these responses were effectively alleviated by exogenous treatment with MT. Exogenous MT reduces oxidative damage and lipid peroxidation by reducing salt-induced ROS and protects the plasma membrane from oxidative toxicity. MT also reduces the osmotic pressure by reducing the salt-induced accumulation of Na⁺ and increasing the contents of K⁺ and proline. Exogenous MT can facilitate stomatal opening and protect the integrity of cotton chloroplast grana lamella structure and mitochondria under salt stress, protect the photosynthetic system of plants, and improve their biomass. An anatomical analysis of leaves and stems showed that MT can improve xylem and phloem and other properties and aides in the transportation of water, inorganic salts, and organic substances. Therefore, the application of MT attenuates salt-induced stress damage to plants. Treatment with exogenous MT positively increased the salt tolerance of cotton seedlings by improving their photosynthetic capacity, stomatal characteristics, ion balance, osmotic substance biosynthetic pathways, and chloroplast and anatomical structures (xylem vessels and phloem vessels). Conclusions: Our study attributes help to protect the structural stability of photosynthetic organs and increase the amount of material accumulation, thereby reducing salt-induced secondary stress. The mechanisms of MT-induced plant tolerance to salt stress provide a theoretical basis for the use of MT to alleviate salt stress caused by unreasonable irrigation, fertilization, and climate change.

Root systems are the key organs through which plants absorb water and nutrients and perceive the soil environment and thus are easily damaged by salt stress. Melatonin can alleviate stress-induced damage to roots. The present study investigated the effects of exogenous melatonin on the root physiology, transcriptome and metabolome of cotton seedlings under salt stress. Salt stress was observed to damage the cell structure and disorder the physiological system of cotton seedling roots. After subjecting melatonin-soaked seeds to salt stress, the activities of SOD, CAT and POD in cotton seedling roots increased by 10–25%, 50–60% and 50–60%, respectively. The accumulation of H₂O₂ and MDA were significantly decreased by 30–60% and 30–50%, respectively. The contents of soluble sugar, soluble protein and K⁺ increased by 15–30%, 15–30% and 20–50%, respectively, while the Na⁺ content was significantly reduced. Melatonin also increased auxin (by 20–40%), brassinosteroids (by 5–40%) and gibberellin (by 5–35%) and promoted melatonin content and root activity. Exogenous melatonin maintained the integrity of root cells and increased the number of organelles. Transcriptomic and metabolomic results showed that exogenous melatonin could mitigate the salt-stress-induced inhibition of plant root development by regulating the reactive oxygen species scavenging system; ABC transporter synthesis; plant hormone signal transduction, endogenous melatonin gene expression; and the expression of the transcription factors MYB, TGA and WRKY33. These results provide a new direction and empirical basis for improving crop salt tolerance with melatonin.

Melatonin is a small-molecule indole hormone that plays an important role in participating in biotic and abiotic stress resistance. Melatonin has been confirmed to promote the normal development of plants under adversity stress by mediating physiological regulation mechanisms. However, the mechanisms by which exogenous melatonin mediates salt tolerance via regulation of antioxidant activity and osmosis in cotton seedlings remain largely unknown. In this study, the regulatory effects of melatonin on reactive oxygen species (ROS), the antioxidant system, and osmotic modulators of cotton seedlings were determined under 0–500 µM melatonin treatments with salt stress induced by 150 mM NaCl treatment. Cotton seedlings under salt stress exhibited an inhibition of growth, excessive hydrogen peroxide (H₂O₂), superoxide anion (O₂⁻), and malondialdehyde (MDA) accumulations in leaves, increased activity levels of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX), and elevated ascorbic acid (AsA) and glutathione (GSH) content in leaves. However, the content of osmotic regulators (i.e., soluble sugars and proteins) in leaves was reduced under salt stress. This indicates high levels of ROS were produced, and the cell membrane was damaged. Additionally, osmotic regulatory substance content was reduced, resulting in osmotic stress, which seriously affected cotton seedling growth under salt stress. However, exogenous melatonin at different concentrations reduced the contents of H₂O₂, O₂⁻, and MDA in cotton leaves, increased the activity of antioxidant enzymes and the content of reductive substances (i.e., AsA and GSH), and promoted the accumulation of osmotic regulatory substances in leaves under salt stress. These results suggest that melatonin can inhibit ROS production in cotton seedlings, improve the activity of the antioxidant enzyme system, raise the content of osmotic regulation substances, reduce the level of membrane lipid peroxidation, and protect the integrity of the lipid membrane under salt stress, which reduces damage caused by salt stress to seedlings and effectively enhances inhibition of salt stress on cotton seedling growth. These results indicate that 200 µM melatonin treatment has the best effect on the growth and salt tolerance of cotton seedlings.

The objective of this study was to assess the impacts of nitrogen on the physiological characteristics of the source–sink system of upper fruiting branches under various amounts of nitrogen fertilization. A two-year field experiment was conducted with a Bt cotton cultivar in the Yellow River Basin of China. The growth and yield of cotton of the upper fruiting branches were compared under four nitrogen levels: Control (N0, 0 kg ha⁻¹), low nitrogen (N1, 120 kg ha⁻¹), moderate nitrogen (N2, 240 kg ha⁻¹), and high nitrogen (N3, 480 kg ha⁻¹). The results indicated that in the subtending leaves in upper fruiting branches, chlorophyll content, protein content, and peroxidase (POD) activity dramatically increased with nitrogen application, reaching the highest under the moderate nitrogen treatment. The physiological characters in the seeds had the same trends as in the subtending leaves. Furthermore, the moderate nitrogen rate (240 kg ha⁻¹) had a favorable yield and quality. Our results supported that a moderate nitrogen rate (240 kg ha⁻¹) could coordinate the source–sink growth of cotton in the late stage, enhance the yield and fiber quality, and decrease the cost of fertilizer in the Yellow River Basin of China and other similar ecological areas.