Seed germination is of great significance for plant development and crop yield. Recently, nitric oxide (NO) has been shown to not only serve as an important nitrogen source during seed development but also to participate in a variety of stress responses in plants to high salt, drought, and high temperature. In addition, NO can affect the process of seed germination by integrating multiple signaling pathways. However, due to the instability of NO gas activity, the network mechanism for its fine regulation of seed germination remains unclear. Therefore, this review aims to summarize the complex anabolic processes of NO in plants, to analyze the interaction mechanisms between NO-triggered signaling pathways and different plant hormones such as abscisic acid (ABA) and gibberellic acid (GA), ethylene (ET) and reactive oxygen species (ROS) signaling molecules, and to discuss the physiological responses and molecular mechanisms of seeds during the involvement of NO in abiotic stress, so as to provide a reference for solving the problems of seed dormancy release and improving plant stress tolerance.

The induction and proliferation of embryogenic callus are key steps for large-scale propagation of somatic embryogenesis pathway and long-term preservation of coniferous germplasm. Callus can be induced from immature embryos of Korean pine (Pinus koraiensis Sieb. et Zucc.; Pinaceae) as explants, but there are problems, such as low proliferation efficiency, loss of embryogenicity, poor vigor; thus, best conditions for proliferation and culture of immature embryos of Korean pine are not yet clear. To solve the problems with somatic embryogenesis of Korean pine and determine the best culture conditions for callus induction and proliferation, we varied hormone concentration, subculture cycle of proliferation and other plant growth regulators combinations in media to induce callus formation by megagametophytes of three Korean pine families at different developmental stages, then analyzed the effects on embryogenic callus retention and cell proliferation using a quadratic regression orthogonal rotation design. The results showed that the family origin and collection date of explants significantly affected callus induction (induction rate reached 1.67%). Embryogenic maintenance and callus proliferation were best on DCR medium supplemented with 0.25 mg L⁻¹ 6-benzyl adenine, 1 mg L⁻¹ naphthaleneacetic acid, 30 g L⁻¹ sucrose, 500 mg L⁻¹, l-glutamine, 500 mg L⁻¹ casein hydrolysis and 6.5 g L⁻¹ agar. In addition, the combination of 2,4-dichlorophenoxyacetic acid + 6-benzyl adenine also had a better proliferative effect on callus. The effects of different combinations of growth regulators on callus proliferation efficiency were significantly different. Transfer to new medium every 13–15 days not only maintained robust callus vigor, but also yielded a larger proliferation coefficient. The techniques and conditions for embryogenic callus induction and proliferation of Korean determined here will serve as a foundation for establishing a large-scale system for somatic embryogenesis and propagation of Korean pine.

Korean pine broadleaf mixed forest is an important ecosystem for maintaining biodiversity in Northeast China. Korean pine is also an important species for the production of timber and nuts in the mountainous areas of Northeast China. In this study, we compared three types of Korean pine callus and found that embryogenic callus had high amounts of storage substances (protein, sugar and starch). Non-embryonic callus had high levels of polyphenols and polyphenol oxidation, while callus that lost somatic embryogenesis potential had lower levels of storage substances (protein, sugar and starch) and higher contents of peroxidase and catalase. These results indicate that high contents of storage substances (protein, sugar and starch), and low levels of polyphenols and polyphenol oxidase can be used as physiological markers of callus with somatic embryogenic potential. During the development process of Korean pine somatic embryos, fresh weight and dry weight gradually increased, while water content gradually decreased. Soluble protein, starch, soluble sugar and superoxide dismutase also increased during development, while peroxidase and catalase levels reduced over time. These results indicate that somatic embryogenesis involves energy storage, and antioxidant enzymes cooperate to regulate the occurrence and development of embryos. These results provide physiological markers for identification of embryogenic callus with somatic embryogenesis, to evaluate callus suitable for somatic embryogenesis, and provide basis for further research on the molecular mechanisms of somatic embryogenesis.

Manchurian ash (Fraxinus mandshurica Rupr.) is a valuable hardwood species in Northeast China. In cultures of F. mandshurica, somatic embryos were produced mainly on browned explants. Therefore, we studied the mechanism of explant browning and its relationship with somatic embryogenesis (SE). We used explants derived from F. mandshurica immature zygotic embryo cotyledons as materials. Proteins were extracted from browned embryogenic explants, browned non-embryogenic explants, and non-brown explants, and then separated by 2-dimensional electrophoresis. Differentially and specifically expressed proteins were analyzed by mass spectrometry to identify proteins involved in the browning of explants and SE. Some stress response and defense proteins such as chitinases, peroxidases, aspartic proteinases, and an osmotin-like protein played important roles during SE of F. mandshurica. Our results indicated that explant browning might not be caused by the accumulation and oxidation of polyphenols only, but also by some stress-related processes, which were involved in programmed cell death (PCD), and then induced SE.