Flooding is the main cause of inhibited seed germination and seedling growth during the direct seeding of rice. As a growth-regulating hormone, melatonin plays an important role in promoting seed germination and enhancing plant stress resistance. However, there are still many unknown mechanisms related to the exogenous-melatonin-mediated regulation of submergence tolerance during seed germination. To overcome these challenges, this study investigated the effects of seed-soaking with different melatonin concentrations on the germination and physiological characteristics of rice seeds over different flooding times. The results showed that the exogenous administration of 100 μM of melatonin significantly improved the germination potential, germination rate, germination index, and vigor index of seeds and increased the size of the aboveground and underground biomass. By increasing the activities of superoxide dismutase (SOD) and peroxidase (POD) in shoots and roots, and reducing the content of malondialdehyde (MDA), the 100 μM melatonin alleviated the damage of rice seeds under flooding stress. The gray relational grade between the root MDA content and the melatonin seed-soaking treatment was the highest, and this was used to evaluate the effect of melatonin in alleviating flooding stress. A two-factor analysis of variance showed that the flooding stress treatment and the melatonin treatment had more significant effects on rice seed germination than the flooding and melatonin interaction treatments (p < 0.01). The optimal number of days for rice seeds to resist flooding stress was two, and the optimal concentration for the melatonin seed-soaking treatment was 100 μM. In sum, a 100 μM melatonin seed-soaking treatment can effectively alleviate the damage of flooding stress on rice seeds and promote the growth and development of seedlings. This study provides the theoretical basis for a light and simple cultivation technique for the direct seeding of rice.

Rice is an important food crop. Rice seedlings are mainly composed of root, coleoptile, mesocotyl and euphylla. The elongation of coleoptile and mesocotyl promotes the emergence of rice seedlings. Therefore, analyzing the mechanism of coleoptile and mesocotyl elongation is important for the cultivation of rice varieties. Due to global warming, heat stress is threatening rice yields. Betaine plays an important role in plant resistance to heat stress; however, we lack research on its regulation mechanism of rice seed germination under heat stress. Therefore, we explored the effects of soaking seeds with betaine at different concentrations on rice seed germination under heat stress. According to the results, soaking seeds with 10 mM of betaine could effectively improve the seeds’ germination potential and rate under heat stress to promote the germination of rice seeds. To clarify the mitigation mechanism of betaine in heat stress, we measured the antioxidant enzyme activity, malondialdehyde content, soluble protein content and endogenous hormone content of seed protrusion under heat stress. We constructed the cDNA library for transcriptome sequencing. According to the results, 10 mM of betaine improved the activities of the superoxide dismutase, peroxidase and catalase of seed protrusion under heat stress to reduce the malondialdehyde content and increase the soluble protein content to alleviate the effect of heat stress on rice seed germination. The detection of the endogenous hormone content showed that soaking seeds with 10 mM of betaine increased the content of gibberellin and decreased the contents of auxin and abscisic acid of seed protrusion under heat stress. According to the transcriptome analysis, betaine can induce the expressions of key genes in the biosynthesis and metabolism of auxin, abscisic acid and gibberellins in the seed coleoptile and mesocotyl elongation stage, regulate the signal transduction of three hormones and promote the germination of rice seeds under heat stress. This study revealed, for the first time, the physiological and molecular regulation mechanism of betaine promotion of seed germination under heat stress.

Drought is a severe abiotic stress that imposes a serious threat to rice productivity. Although previous studies have found that melatonin can promote seed germination, the physiological regulation mechanism of drought tolerance in rice seed germination mediated by exogenous melatonin is still unclear. In order to overcome these challenges, polyethylene glycol 6000 (PEG6000) at concentrations of 20% and 35% was used to simulate osmotic stress. Rice seeds were treated with different concentrations of melatonin (i.e., 0, 20, 100, and 500 μM) to study the effects on germination characteristics, growth and development, superoxide dismutase (SOD) activity, peroxidase (POD) activity, catalase (CAT) activity, malondialdehyde (MDA) content, and soluble protein content. The results showed that the seed soaking treatment using melatonin at a concentration of 100 μM under drought stress effectively promoted the germination rate and improved the biomass of rice seed shoots and roots. Meanwhile, this treatment reduced MDA content to alleviate the oxidative damage of rice seeds caused by drought stress. The two-way ANOVA showed that the effect of single melatonin soaking treatment on rice seed germination was more significant than that of single drought stress and the interaction of drought stress and melatonin (p < 0.05). Using the membership function method, it was shown that the critical gradient of rice seeds under drought stress was 35%, and the critical treatment of interactive treatment was 35% + 100 μM. Through grey correlation analysis, it was found that germination rate (7 d) had the highest grey correlation with melatonin seed soaking treatment to evaluate the mitigation effect of melatonin on drought stress. This study provides a theoretical basis for light and simple cultivation technology for the dry direct seeding of rice.