Understanding the relationship between exogenous melatonin and water deficit stress is crucial for achieving high yields and alleviating the effects of water deficit stress on soybean (Glycine max (L.) Merrill) plants in agriculture. This study investigated the effects of exogenous melatonin on soybean photosynthetic capacity under water deficit stress induced by polyethylene glycol (PEG) 6000. We conducted a potting experiment in 2018 using the soybean (Glycine max L. Merrill) cultivar Suinong 26. We identified the impacts of a concentration of PEG 6000 simulating drought (15%, w/v) and an appropriate melatonin concentration (100 μmol/L) on the growth of soybean seedlings and flowering stages in a preliminary test. We applied exogenous melatonin by foliar spraying and root application to determine the effects on leaf photosynthesis during water deficit stress. Our results indicated that 15% PEG 6000 had an obvious inhibitory effect on the growth of soybean seedlings and flowering stages, causing oxidative stress and damage due to reactive oxygen species (ROS) (H₂O₂ and O₂^·-) accumulation and potentially reducing air exchange parameters and photosystem II (PSII) efficiency. The application of exogenous melatonin significantly relieved the inhibitory effects of PEG 6000 stress on seedlings and flowering growth, and gas exchange parameters, potentially improved PSII efficiency, improved the leaf area index (LAI) and the accumulation of dry matter, slowed down oxidative stress and damage to leaves by increasing the activity of antioxidant enzymes (SOD, POD, and CAT), reduced the content of malondialdehyde (MDA), and ultimately improved soybean yield. Overall, the results of this study demonstrated that application of exogenous melatonin at the seedlings and flowering stages of soybean is effective in alleviating plant damage caused by water deficit stress and improving the drought resistance of soybean plants. In addition, the results showed that application of exogenous melatonin by root is superior to foliar spraying.

Introduction: Melatonin is a multipotent molecule that exists widely in animals and plants and plays an active regulatory role in abiotic stresses. The B3 superfamily is a ubiquitous transcription factor with a B3 functional domain in plants, which can respond temporally to abiotic stresses by activating defense compounds and plant hormones. Despite the fact that the B3 genes have been studied in a variety of plants, their role in soybean is still unknown.Methods: The regulation of melatonin on cold resistance of soybean and the response of B3 genes to cold stress were investigated by measuring biochemical indexes of soybean. Meanwhile, the genome-wide identification of B3 gene family was conducted in soybean, and B3 genes were analyzed based on phylogeny, motifs, gene structure, collinearity, and cis-regulatory elements analysis.Results: We found that cold stress-induced oxidative stress in soybean by producing excessive reactive oxygen species. However, exogenous melatonin treatment could increase the content of endogenous melatonin and other hormones, including IAA and ABA, and enhance the antioxidative system, such as POD activity, CAT activity, and GSH/GSSG, to scavenge ROS. Furthermore, the present study first revealed that melatonin could alleviate the response of soybean to cold stress by inducing the expression of B3 genes. In addition, we first identified 145 B3 genes in soybean that were unevenly distributed on 20 chromosomes. The B3 gene family was divided into 4 subgroups based on the phylogeny tree constructed with protein sequence and a variety of plant hormones and stress response cis-elements were discovered in the promoter region of the B3 genes, indicating that the B3 genes were involved in several aspects of the soybean stress response. Transcriptome analysis and results of qRT-PCR revealed that most GmB3 genes could be induced by cold, the expression of which was also regulated by melatonin. We also found that B3 genes responded to cold stress in plants by interacting with other transcription factors.Discussion: We found that melatonin regulates the response of soybean to cold stress by regulating the expression of the transcription factor B3 gene, and we identified 145 B3 genes in soybean. These findings further elucidate the potential role of the B3 gene family in soybean to resist low-temperature stress and provide valuable information for soybean functional genomics study.

Soda saline-alkali soil has double adverse effects on growth, morphogenesis and yield of the crops by creating a salinity stress and a high pH value in the rhizosphere surroundings of the plant. Seed priming can be an effective approach to enhance stress adaptation in seedlings growing in salinity stressed conditions. The present study examined the role of hydropriming and comprehensive seed priming with ZnSO₄, CaCl₂, betaine hydrochloride and GA₃ to enhance the soda saline-alkali tolerance in soybeans (Glycine max [L.] Merr.) seedlings. An unprimed control treatment was also maintained for comparison. The results show that two seed priming treatments can effectively mitigate the negative effects of soda saline-alkali stress. Nevertheless, compared with hydropriming, soybean seedlings from the comprehensive seed priming treatment had better growth performance through accumulation of soluble sugar and free proline contents, enhancing the activities of superoxide dismutase (SOD) and catalase (CAT), decreasing the malondialdehyde (MDA) content formed from lipid peroxidation, increasing the photosynthetic pigment contents, maintaining a better membrane integrity of the chloroplasts and mitochondria, and generating more starch grains in the chloroplasts of spongy mesophyll cells under soda saline-alkali stress. In conclusion, our results suggest that soybean seedlings from comprehensive seed priming exhibited a higher tolerance to soda saline-alkali stress than the unprimed control treatment.

There is a growing need to enhance the productivity of soybean (Glycine max L.) under severe drought conditions in order to improve global food security status. Melatonin, a ubiquitous hormone, could alleviate drought stress in various plants. Earlier, we demonstrated that exogenous melatonin treatment could enhance the tolerance of drought-treated soybean. However, the underlying mechanisms by which this hormone exerts drought resistance is still unclear. The present study used transcriptomic and metabolomic techniques to determine some critical genes and pathways regulating melatonin response to drought conditions. Results showed that exogenous melatonin treatment could increase relative water content and decrease electrolyte leakage in the leaves and increase seed yield under drought stress. Transcriptomic analysis showed that there were 852 core differentially expressed genes (DEGs) that were regulated by drought stress and melatonin in soybean leaves. The most enriched drought-responsive genes are mainly involved in the ‘biosynthesis of secondary metabolites’. Metabolomic profiling under drought stress showed higher accumulation levels of secondary metabolites related to drought tolerance after exogenous melatonin treatment. Also, we highlighted the vital role of the pathways including phenylpropanoid, flavonoid, isoflavonoid, and steroid biosynthesis pathways for improvement of drought tolerance in soybean by exogenous melatonin treatment. In all, findings from this study give detailed molecular basis for the application of melatonin as a drought-resistant agent in soybean cultivation.